Methods for treating hypercholesterolemia

ABSTRACT

Disclosed herein are antisense compounds and methods for decreasing LDL-C in an individual having elevated LDL-C. Additionally disclosed are antisense compounds and methods for treating, preventing, or ameliorating hypercholesterolemia and/or atherosclerosis. Further disclosed are antisense compounds and methods for decreasing coronary heart disease risk. Such methods include administering to an individual in need of treatment an antisense compound targeted to a PCSK9 nucleic acid. The antisense compounds administered include gapmer antisense oligonucleotides.

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing is provided as a file entitledBIOL0090USC4SEQ_ST25.txt, created on Apr. 6, 2017 which is 144 Kb insize. The information in the electronic format of the sequence listingis incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

Methods are provided for lowering LDL-C levels in an individual havingelevated LDL-C levels. Such methods are further useful to treathypercholesterolemia, and to reduce the risk of coronary heart disease.

BACKGROUND

Atherosclerosis is a complex, polygenic disease of arterial degenerationthat can lead to coronary heart disease (CHD). In Western societies,complications arising from atherosclerosis are the most common causes ofdeath. Several risk factors for CHD have been well-established, andinclude elevated low density lipoprotein-cholesterol (LDL-C), low levelsof high density lipoprotein cholesterol (HDL-C), cigarette smoking,hypertension, age, and family history of early CHD. Given the abundantclinical data and epidemiological studies indicating that lowering LDL-Cis beneficial for the prevention of adverse coronary events, the primarytarget of pharmacological intervention in the treatment and preventionof CHD is lowered LDL-C.

In individuals with autosomal dominant hypercholesterolemia (ADH),elevated LDL-C levels have been linked to mutations in the genesencoding LDL-receptor (LDL-R), apolipoprotein B (apoB), or proproteinconvertase subtilisin/kexin type 9 (PCSK9) (Abifadel et al., Nat.Genet., 2003, 34:154-156). The PCSK9 gene (also known as FH3; NARC1;NARC-1; and HCHOLA3) was mapped to Chromosome 1 at location 1p32.3.PCSK9 was identified as a third locus associated with ADH whengain-of-function mutations in PCSK9 were found to be linked to elevatedLDL-C levels. ApoB-100 participates in the intracellular assembly andsecretion of triglyceride-rich lipoproteins and is a ligand for theLDL-R. PCSK9 is proposed to reduce LDL-R expression levels in the liver.Reduced LDL-R expression results in reduced hepatic uptake ofcirculating apoB-100-containing lipoproteins, which in turn leads toelevated cholesterol.

Familial hypercholesterolemia (FH) is caused by hundreds of differentmutations in the LDL-R, and is phenotypically characterized by elevatedplasma LDL-C levels and deposits of LDL-C in tendons, skin and arteries,leading to premature cardiovascular disease. Homozygous and heterozygousmutations in the LDL-R are associated with FH. Likewise, heterozygousand homozygous mutations in the ligand-binding domain of PCSK9 areassociated with the FH phenotype. Mutations in this gene have beenassociated with a third form of autosomal dominant FH.

In mice genetically deficient in PCSK9, a marked increase in LDL-Rexpression and increased plasma LDL clearance rate were observed.Conversely, overexpression in mice of wild-type PCSK9, or certain PCSK9mutants, promotes decreased LDL-R expression and elevated LDL-C levels.

SUMMARY OF THE INVENTION

Provided herein are methods for the treatment of hypercholesterolemiaand/or atherosclerosis, as well as methods for the reduction of elevatedLDL-C levels and CHD risk. Methods for treating hypercholesterolemiacomprise administering to an individual an antisense compound targetedto a PCSK9 nucleic acid. In such methods, an antisense compound targetedto a PCSK9 nucleic acid may be targeted to sequences as set forth inGENBANK® Accession No. NM_174936.2, nucleotides 25475000 to 25504000 ofGENBANK® Accession No. NT_032977.8, or GENBANK® Accession No.AK124635.1.

Methods for treating and/or preventing atherosclerosis compriseadministering to an individual an antisense compound targeted to a PCSK9nucleic acid. Methods for reducing LDL-C levels comprise administeringto an individual an antisense compound targeted to a PCSK9 nucleic acid.Methods for reducing CHD risk comprise administering to an individual anantisense compound targeted to a PCSK9 nucleic acid. Such methods maycomprise the administration of a therapeutically effective amount of anantisense compound targeted to a PCSK9 nucleic acid.

Methods are provided for reducing LDL-C levels in an individual havingelevated LDL-C levels, comprising administering to the individual atherapeutically effective amount of an antisense oligonucleotidetargeted to a PCSK9 nucleic acid, thereby reducing LDL-C levels. Alsoprovided are methods for reducing LDL-C levels in an individualcomprising selecting an individual having elevated LDL-C levels, andadministering to the individual a therapeutically effective amount of anantisense compound targeted to a PCSK9 nucleic acid, and additionallymonitoring LDL-cholesterol levels. Further provided are methods fortreating atherosclerosis in an individual, comprising selecting anindividual diagnosed with atherosclerosis, administering to theindividual a therapeutically effective amount of an antisense compoundtargeted to a PCSK9 nucleic acid, and monitoring atherosclerosis. Alsoprovided are methods for reducing coronary heart disease risk,comprising selecting an individual having elevated LDL-C levels and oneor more additional indicators of coronary heart disease, administeringto the individual a therapeutically effective amount of an antisensecompound targeted to a PCSK9 nucleic acid, and monitoring LDL-C levels.Additionally provided are methods for treating hypercholesterolemia,comprising administering to an individual diagnosed withhypercholesterolemia a therapeutically effective amount of an antisenseoligonucleotide targeted to a PCSK9 nucleic acid, thereby reducingcholesterol levels.

Further provided is a use of an effective amount of one or more of thedisclosed antisense compounds for treating atherosclerosis orhypercholesterolemia in an individual in need of such treatment. Alsoprovided is a use of an effective amount of one or more of the disclosedantisense compounds for reducing LDL-C levels or reducing CHD risk in anindividual in need of such treatment. Also provided is the use of one ormore of the disclosed antisense compounds in the manufacture of amedicament for the treatment of atherosclerosis or hypercholesterolemia.Further provided is the use of one or more of the disclosed antisensecompounds in the manufacture of a medicament for reducing LDL-C levelsor for reducing CHD risk.

In any of the methods provided, a PCSK9 nucleic acid may be the sequenceset forth in SEQ ID NO: 1. Thus, the antisense compound may be targetedto a PCSK9 nucleic acid as set forth in SEQ ID NO: 1.

Any of the methods provided herein may further comprise monitoring LDL-Clevels.

An individual may be selected for administration of an antisensecompound targeted to a PCSK9 nucleic acid when the individual exhibitsan LDL-C level above 100 mg/dL, above 130 mg/dL, above 160 mg/dL, orabove 190 mg/dL. Administration of an antisense compound targeted to aPCSK9 nucleic acid may result in an LDL-C level below 190 mg/dL, below160 mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50mg/dL.

In any of the aforementioned methods, administration of the antisensecompound may comprise parenteral administration. The parenteraladministration may further comprise subcutaneous or intravenousadministration.

In any of the methods provided herein, the antisense compound may haveleast 80%, at least 90%, or at least 95% complementarity to SEQ IDNO: 1. Alternatively, the antisense compound may have 100%complementarity to SEQ ID NO: 1.

The antisense compounds provided herein and employed in any of thedescribed methods may be 8 to 80 subunits in length, 12 to 50 subunitsin length, 12 to 30 subunits in length, 15 to 30 subunits in length, 18to 24 subunits in length, 19 to 22 subunits in length, or 20 subunits inlength. Further, the antisense compounds employed in any of thedescribed methods may be antisense oligonucleotides 8 to 80 nucleotidesin length, 12 to 50 nucleotides in length, 12 to 30 nucleotides inlength 15 to 30 nucleotides in length, 18 to 24 nucleotides in length,19 to 22 nucleotides in length, or 20 nucleotides in length.

In any of the methods provided, the antisense compound may be anantisense oligonucleotide. Moreover, the antisense oligonucleotide maybe a gapmer antisense oligonucleotide. The gapmer antisenseoligonucleotide may comprise a gap segment of ten 2′-deoxynucleotidespositioned between wing segments of five 2′-MOE nucleotides. The gapmerantisense oligonucleotide may be a gap-widened antisenseoligonucleotide, comprising a gap segment of fourteen2′-deoxynucleotides positioned between wing segments of three 2′-MOEnucleotides.

In any of the methods provided, the antisense compounds may have atleast one modified internucleoside linkage. Additionally, eachinternucleoside linkage may be a phosphorothioate internucleosidelinkage. Each cytosine may be a 5-methyl cytosine.

Also provided herein are antisense compounds targeted to a PCSK9 nucleicacid. Further provided are antisense oligonucleotides targeted to aPCSK9 nucleic acid. The antisense compounds, including antisenseoligonucleotides, may be targeted to PCSK9 nucleic acids, which includethe sequences as set forth in GENBANK® Accession No. NM_174936.2,nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_032977.8,or GENBANK® Accession No. AK124635.1. The antisense compounds, includingantisense oligonucleotides, may have at least 70%, at least 80%, atleast 90%, or at least 95% complementarity to a PCSK9 nucleic acid. Theantisense compounds, including antisense oligonucleotides, may have 99%complementarity to a PCSK9 nucleic acid. The antisense compounds,including antisense oligonucleotides, may have 100% complementarity to aPCSK9 nucleic acid. For any of the antisense compounds provided,including antisense oligonucleotides, the PCSK9 nucleic acid may be thesequence set forth in SEQ ID NO: 1.

Antisense compounds targeted to a PCSK9 nucleic acid may be 8 to 80subunits in length, 12 to 50 subunits in length, 12 to 30 subunits inlength, 15 to 30 subunits in length, 18 to 24 subunits in length, 19 to22 subunits in length, or 20 subunits in length. Antisenseoligonucleotides targeted to a PCSK9 nucleic acid may be 8 to 80nucleotides in length, 12 to 50 nucleotides in length, 12 to 30nucleotides in length, 15 to 30 nucleotides in length, 18 to 24nucleotides in length, 19 to 22 nucleotides in length, or 20 nucleotidesin length.

Moreover, the antisense compound may be a gapmer antisenseoligonucleotide. The gapmer antisense oligonucleotide may comprise a gapsegment of ten 2′-deoxynucleotides positioned between wing segments offive 2′-MOE nucleotides. The gapmer antisense oligonucleotide may be agap-widened antisense oligonucleotide, comprising a gap segment offourteen 2′-deoxynucleotides positioned between wing segments of three2′-MOE nucleotides.

Antisense compounds, including antisense oligonucleotides, targeted to aPCSK9 nucleic acid may have at least one modified internucleosidelinkage. Additionally, each internucleoside linkage may be aphosphorothioate internucleoside linkage. Each cytosine may be a5-methyl cytosine.

Antisense compounds, including antisense oligonucleotides, targeted to aPCSK9 nucleic acid may have at least one modified nucleoside. In certainembodiments, a modified nucleoside is a sugar-modified nucleoside. Incertain such embodiments, the sugar-modified nucleosides can furthercomprise a natural or modified heterocyclic base moiety and/or a naturalor modified internucleoside linkage and may include furthermodifications independent from the sugar modification. In certainembodiments, a sugar modified nucleoside is a 2′-modified nucleoside,wherein the sugar ring is modified at the 2′ carbon from natural riboseor 2′-deoxy-ribose.

In certain embodiments, a 2′ modified nucleoside has a 2′-F, 2′-OCH₂(2′-OMe) or a 2′-O(CH₂)₂—OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituentgroup

In certain embodiments, a 2′-modified nucleoside has a bicyclic sugarmoiety. In certain such embodiments, the bicyclic sugar moiety is a Dsugar in the alpha configuration. In certain such embodiments, thebicyclic sugar moiety is a D sugar in the beta configuration. In certainsuch embodiments, the bicyclic sugar moiety is an L sugar in the alphaconfiguration. In certain such embodiments, the bicyclic sugar moiety isan L sugar in the beta configuration.

In certain embodiments, the bicyclic sugar moiety comprises a bridgegroup between the 2′ and the 4′-carbon atoms. In certain suchembodiments, the bridge group comprises from 1 to 8 linked biradicalgroups. In certain embodiments, the bicyclic sugar moiety comprises from1 to 4 linked biradical groups. In certain embodiments, the bicyclicsugar moiety comprises 2 or 3 linked biradical groups. In certainembodiments, the bicyclic sugar moiety comprises 2 linked biradicalgroups. In certain embodiments, a linked biradical group is selectedfrom —O—, —S—, —N(R1)-, —C(R1)(R2)-, —C(R1)=C(R1)-, —C(R1)=N—,—C(═NR1)-, —Si(R1)(R2)-, —S(═O)2-, —S(═O)—, —C(═O)— and —C(═S)—; whereeach R1 and R2 is, independently, H, hydroxyl, C1-C12 alkyl, substitutedC1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20aryl, a heterocycle radical, a substituted hetero-cycle radical,heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substitutedC5-C7 alicyclic radical, halogen, substituted oxy (—O—), amino,substituted amino, azido, carboxyl, substituted carboxyl, acyl,substituted acyl, CN, thiol, substituted thiol, sulfonyl (S(═O)2-H),substituted sulfonyl, sulfoxyl (S(═O)—H) or substituted sulfoxyl; andeach substituent group is, independently, halogen, C1-C12 alkyl,substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl,C2-C12 alkynyl, substituted C2-C12 alkynyl, amino, substituted amino,acyl, substituted acyl, C1-C12 aminoalkyl, C1-C12 aminoalkoxy,substituted C1-C12 aminoalkyl, substituted C1-C12 aminoalkoxy or aprotecting group.

In some embodiments, the bicyclic sugar moiety is bridged between the 2′and 4′ carbon atoms with a biradical group selected from —O—(CH2)p-,—O—CH2-, —O—CH2CH2-, —O—CH(alkyl)-, —NH—(CH2)p-, —N(alkyl)-(CH2)p-,—O—CH(alkyl)-, —(CH(alkyl))-(CH2)p-, —NH—O—(CH2)p-, —N(alkyl)-O—(CH2)p-,or —O—N(alkyl)-(CH2)p-, wherein p is 1, 2, 3, 4 or 5 and each alkylgroup can be further substituted. In certain embodiments, p is 1, 2 or3.

In one aspect, each of said bridges is, independently, —[C(R1)(R2)]n-,—[C(R1)(R2)]n-O—, —C(R1R2)-N(R1)-O— or —C(R1R2)-O—N(R1)-. In anotheraspect, each of said bridges is, independently, 4′-(CH2)3-2′,4′-(CH2)2-2′, 4′-CH2-O-2′, 4′-(CH2)2-O-2′, 4′-CH2-O—N(R1)-2′ and4′-CH2-N(R1)-O-2′- wherein each R1 is, independently, H, a protectinggroup or C1-C12 alkyl.

The present disclosure further discloses subsets of the antisensecompounds above that show superior properties relating topharmacodynamics and/or pharmacokinetics, among other properties.Exemplary antisense compounds reduce PCSK9 expression in a cultured cellmodel system, e.g., Hep3B cells, cynomolgus monkey hepatocytes, or humanprimary hepatocytes. In particular embodiments, antisense compoundsreduce PCSK9 expression in a dose-dependent manner in a cell culturesystem, where the antisense compounds have an IC₅₀ in the nanomolarrange.

Exemplary antisense compounds also reduce the expression of a PCSK9nucleic acid in an animal model, such as a transgenic mouse thatexpresses a human PCSK9 nucleic acid (HuPCSK9Tg mice). In particularembodiments, antisense compounds include those that reduce PCSK9 expressin an animal model with a physiology that correlates closely withhumans, such as monkey, e.g., the cynomolgus monkey.

Exemplary antisense compounds display minimal side effects. Side effectsinclude responses to the administration of the antisense compoundunrelated to the targeting of a PCSK9 nucleic acid, such as aninflammatory response in the host individual. Exemplary antisensecompounds are well tolerated by the host individual. Tolerability may bedetermined though histological analysis of various cell types, andincludes examination of such markers as cytoplasmic swelling frommultifocal apoptosis in liver cells, follicular hyperplasia in spleencells, macrophage infiltration, and the like. In particular embodiments,antisense compounds produce minimal signs of host intolerance.

Exemplary antisense compounds further display favorablepharmacokinetics. In particular embodiments, antisense compoundsaccumulate at a relatively high ratio in the liver versus othersensitive organs, such as kidneys. In particular embodiments, antisensecompounds exhibit relatively high half-lives in relevant biologicalfluids or tissues, e.g., liver tissue, reflecting higher stability andresistance to nucleases, for example.

In particular embodiments, antisense compounds target identicalsequences in human and animal PCSK9 sequences. In other embodiments,antisense compounds target PCSK9 sequences without a known singlenucleotide polymorphism (SNP), have a high relative G-content, haveminimal secondary structure, have greatly reduced effects on the host'sserum chemistry, body weight, or histopathology, compared to otherantisense compounds, and/or induce a minimal adverseimmunohistocompatibility reaction.

In a particular embodiment, the antisense compound is selected from thegroup of Isis 405881, Isis 399819, Isis 395165, Isis 405879, Isis406008, Isis 405891, Isis 395186, Isis 405988, Isis 405994, Isis 406023,Isis 395187, Isis 395185, Isis 406033, Isis 405923, Isis 399900, Isis405995, Isis 405991, Isis 406005, Isis 399793, and Isis 395152. Theaforementioned antisense compounds effectively repress PCSK9 expressionin a Hep3B cell culture system. In another embodiment, antisensecompounds effectively inhibit PCSK9 expression with a low IC₅₀ in abattery of cell culture systems. Exemplary antisense compounds includethose selected from the group of Isis 395165, Isis 395185, Isis 395186,Isis 395187, Isis 405879, Isis 405881, Isis 405891, Isis 405988, Isis405994, and Isis 406008. In yet another embodiment, antisense compoundsexhibit minimal or no adverse histopathological effects whenadministered at particularly high doses to an individual. Exemplaryantisense compounds include those selected from the group consisting ofIsis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405891, andIsis 405988. In yet another embodiment, antisense compounds affecttherapeutic end-points, e.g., reduction of plasma LDL-C, liver TG, orliver PCSK9 mRNA, in an animal model. Exemplary antisense compoundsinclude Isis 405879. In yet another embodiment, antisense compoundsdisplay combinations of the characteristics above and reduce liver PCSK9mRNA expression in an animal model with high efficiency, e.g., Isis405879.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by on of skill in theart to which the invention(s) belong. Unless specific definitions areprovided, the nomenclature utilized in connection with, and theprocedures and techniques of, analytical chemistry, synthetic organicchemistry, and medicinal and pharmaceutical chemistry described hereinare those well known and commonly used in the art. Standard techniquesmay be used for chemical synthesis, chemical analysis, pharmaceuticalpreparation, formulation and delivery, and treatment of subjects.Certain such techniques and procedures may be found for example in“Antisense Drug Technology: Principles, Strategies, and Applications.”by Stanley Crooke, Boca Raton: Taylor & Francis Group, 2008;“Carbohydrate Modifications in Antisense Research” Edited by Sangvi andCook, American Chemical Society, Washington D.C., 1994; and “Remington'sPharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., 18thedition, 1990; and which is hereby incorporated by reference for anypurpose. Where permitted, all patents, patent applications, publishedapplications and publications, GENBANK sequences, websites and otherpublished materials referred to throughout the entire disclosure herein,unless noted otherwise, are incorporated by reference in their entirety.All of the GENBANK® Accession Nos. along with their associated sequenceand structural data pertaining to such sequences including geneorganization and structural elements and SNP information that may befound in sequence databases such as the National Center forBiotechnology Information (NCBI) are incorporated herein by reference intheir entirety. In the event that there is a plurality of definitionsfor terms herein, those in this section prevail. Where reference is madeto a URL or other such identifier or address, it is understood that suchidentifiers can change and particular information on the internet cancome and go, but equivalent information can be found by searching theinternet. Reference thereto evidences the availability and publicdissemination of such information.

A “pharmaceutical composition” means a mixture of substances suitablefor administering to an individual. For example, a pharmaceuticalcomposition may comprise an antisense oligonucleotide and a sterileaqueous solution.

“Administering” means providing a pharmaceutical agent to an individual,and includes, but is not limited to administering by a medicalprofessional and self-administering.

“Individual” means a human or non-human animal selected for treatment ortherapy.

“Duration” means the period of time during which an activity or eventcontinues. In certain embodiments, the duration of treatment is theperiod of time during which doses of a pharmaceutical agent areadministered.

“Parenteral administration,” means administration through injection orinfusion. Parenteral administration includes, but is not limited to,subcutaneous administration, intravenous administration, orintramuscular administration.

“Subcutaneous administration” means administration just below the skin.“Intravenous administration” means administration into a vein.

“Dose” means a specified quantity of a pharmaceutical agent provided ina single administration. In certain embodiments, a dose may beadministered in two or more boluses, tablets, or injections. Forexample, in certain embodiments, where subcutaneous administration isdesired, the desired dose requires a volume not easily accommodated by asingle injection. In such embodiments, two or more injections may beused to achieve the desired dose. In certain embodiments, a dose may beadministered in two or more injections to minimize injection sitereaction in a individual.

“Dosage unit” means a form in which a pharmaceutical agent is provided.In certain embodiments, a dosage unit is a vial containing lyophilizedantisense oligonucleotide. In certain embodiments, a dosage unit is avial containing reconstituted antisense oligonucleotide.

“Pharmaceutical agent” means a substance provides a therapeutic benefitwhen administered to a individual. For example, in certain embodiments,an antisense oligonucleotide targeted to PCSK9 is pharmaceutical agent.

“Diluent” means an ingredient in a composition that lackspharmacological activity, but is pharmaceutically necessary ordesirable. For example, in drugs that are injected the diluent may be aliquid, e.g., saline solution.

“Active pharmaceutical ingredient” means the substance in apharmaceutical composition that provides a desired effect.

“Pharmaceutically acceptable carrier” means a medium or diluent thatdoes not interfere with the structure of the oligonucleotide. Certain ofsuch carriers enable pharmaceutical compositions to be formulated as,for example, tablets, pills, dragees, capsules, liquids, gels, syrups,slurries, suspension and lozenges for the oral ingestion by a subject.

“Therapeutically effective amount” means an amount of a pharmaceuticalagent that provides a therapeutic benefit to an individual. In certainembodiments, a therapeutically effective amount of antisense compoundtargeted to a PCSK9 nucleic acid is an amount that decreases LDL-C inthe individual.

“Hypercholesterolemia” means a condition characterized by elevated serumcholesterol.

“Hyperlipidemia” means a condition characterized by elevated serumlipids.

“Hypertriglyceridemia” means a condition characterized by elevated serumtriglyceride levels.

“Non-familial hypercholesterolemia” means a condition characterized byelevated serum cholesterol that is not the result of a single genemutation.

“Polygenic hypercholesterolemia” means a condition characterized byelevated cholesterol that results from the influence of a variety ofgenetic factors. In certain embodiments, polygenic hypercholesterolemiamay be exacerbated by dietary intake of lipids.

“Familial hypercholesterolemia (FH)” means an autosomal dominantmetabolic disorder characterized by a mutation in the LDL-receptor(LDL-R) gene, markedly elevated LDL-C and premature onset ofatherosclerosis. A diagnosis of familial hypercholesterolemia is madewhen an individualmeets one or more of the following criteria: genetictesting confirming 2 mutated LDL-receptor genes; genetic testingconfirming one mutated LDL-receptor gene; document history of untreatedserum LDL-cholesterol greater than 500 mg/dL; tendinous and/or cutaneousxanthoma prior to age 10 years; or, both parents have documentedelevated serum LDL-cholesterol prior to lipid-lowering therapyconsistent with heterozygous familial hypercholesterolemia.

“Homozygous familial hypercholesterolemia” or “HoFH” means a conditioncharacterized by a mutation in both maternal and paternal LDL-R genes.

“Heterozygous familial hypercholesterolemia” or “HeFH” means a conditioncharacterized by a mutation in either the maternal or paternal LDL-Rgene.

“Mixed dyslipidemia” means a condition characterized by elevated serumcholesterol and elevated serum triglycerides.

“Diabetic dyslipidemia” or “Type II diabetes with dyslipidemia” means acondition characterized by Type II diabetes, reduced HDL-C, elevatedserum triglycerides, and elevated small, dense LDL particles.

“CHD risk equivalents,” means indicators of clinical atheroscleroticdisease that confer a high risk for coronary heart disease, and includeclinical coronary heart disease, symptomatic carotid artery disease,peripheral arterial disease, and/or abdominal aortic aneurysm.

“Metabolic syndrome” means a condition characterized by a clustering oflipid and non-lipid cardiovascular risk factors of metabolic origin. Incertain embodiments, metabolic syndrome is identified by the presence ofany 3 of the following factors: waist circumference of greater than 102cm in men or greater than 88 cm in women; serum triglyceride of at least150 mg/dL; HDL-C less than 40 mg/dL in men or less than 50 mg/dL inwomen; blood pressure of at least 130/85 mmHg; and fasting glucose of atleast 110 mg/dL.

“Non-alcoholic fatty liver disease (NAFLD)” means a conditioncharacterized by fatty inflammation of the liver that is not due toexcessive alcohol use (for example, alcohol consumption of over 20g/day). In certain embodiments, NAFLD is related to insulin resistanceand the metabolic syndrome.

“Non-alcoholic steatohepatitis (NASH)” means a condition characterizedby inflammation and the accumulation of fat and fibrous tissue in theliver, that is not due to excessive alcohol use. NASH is an extreme formof NAFLD.

“Major risk factors” mean factors that contribute to a high risk forcoronary heart disease, and include without limitation cigarettesmoking, hypertension, low HDL-C, family history of coronary heartdisease, and age.

“CHD risk factors” mean CHD risk equivalents and major risk factors.

“Coronary heart disease (CHD)” means a narrowing of the small bloodvessels that supply blood and oxygen to the heart, which is often aresult of atherosclerosis.

“Reduced coronary heart disease risk” means a reduction in thelikelihood that an individual will develop coronary heart disease. Incertain embodiments, a reduction in coronary heart disease risk ismeasured by an improvement in one or more CHD risk factors, for example,a decrease in LDL-C levels.

“Atherosclerosis” means a hardening of the arteries affecting large andmedium-sized arteries and is characterized by the presence of fattydeposits. The fatty deposits are called “atheromas” or “plaques,” whichconsist mainly of cholesterol and other fats, calcium and scar tissue,and damage the lining of arteries.

“History of coronary heart disease” means the occurrence of clinicallyevident coronary heart disease in the medical history of an individualor a individual's family member.

“Early onset coronary heart disease” means a diagnosis of coronary heartdisease prior to age 50.

“Statin intolerant individual” means an individual who as a result ofstatin therapy experiences one or more of creatine kinase increases,liver function test abnormalities, muscle aches, or central nervoussystem side effects.

“Efficacy” means the ability to produce a desired effect. For example,efficacy of a lipid-lowering therapy may be reduction in theconcentration of one or more of LDL-C, VLDL-C, IDL-C, non-HDL-C, ApoB,lipoprotein(a), or triglycerides.

“Acceptable safety profile” means a pattern of side effects that iswithin clinically acceptable limits.

“Side effects” means physiological responses attributable to a treatmentother than desired effects. In certain embodiments, side effectsinclude, without limitation, injection site reactions, liver functiontest abnormalities, renal function abnormalities, liver toxicity, renaltoxicity, central nervous system abnormalities, and myopathies. Forexample, increased aminotransferase levels in serum may indicate livertoxicity or liver function abnormality. For example, increased bilirubinmay indicate liver toxicity or liver function abnormality.

“Injection site reaction” means inflammation or abnormal redness of skinat a site of injection in an individual.

“Individual compliance” means adherence to a recommended or prescribedtherapy by an individual.

“Lipid-lowering therapy” means a therapeutic regimen provided to anindividual to reduce one or more lipids in a individual. In certainembodiments, a lipid-lowering therapy is provide to reduce one or moreof ApoB, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDL-C,triglycerides, small dense LDL particles, and Lp(a) in a individual.

“Lipid-lowering agent” means a pharmaceutical agent provided to anindividual to achieve a lowering of lipids in the individual. Forexample, in certain embodiments, a lipid-lowering agent is provided toan individual to reduce one or more of ApoB, LDL-C, total cholesterol,and triglycerides.

“LDL-C target” means an LDL-C level that is desired followinglipid-lowering therapy.

“Comply” means the adherence with a recommended therapy by a individual.

“Recommended therapy” means a therapeutic regimen recommended by amedical professional for the treatment, amelioration, or prevention of adisease.

“Low LDL-receptor activity” means LDL-receptor activity that is notsufficiently high to maintain clinically acceptable levels of LDL-C inthe bloodstream.

“Cardiovascular outcome” means the occurrence of major adversecardiovascular events.

“Improved cardiovascular outcome” means a reduction in the occurrence ofmajor adverse cardiovascular events, or the risk thereof. Examples ofmajor adverse cardiovascular events include, without limitation, death,reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiacarrest, and atrial dysrhythmia.

“Surrogate markers of cardiovascular outcome” means indirect indicatorsof cardiovascular events, or the risk thereof. For example, surrogatemarkers of cardiovascular outcome include carotid intimal mediathickness (CIMT). Another example of a surrogate marker ofcardiovascular outcome includes atheroma size. Atheroma size may bedetermined by intravascular ultrasound (IVUS). Surrogate markers alsoinclude increased HDL-cholesterol, or any combination of the markersabove.

“Increased HDL-C” means an increase in serum HDL-C in an individual overtime.

“Lipid-lowering” means a reduction in one or more serum lipids in anindividual over time.

“Co-administration” means administration of two or more pharmaceuticalagents to a individual. The two or more pharmaceutical agents may be ina single pharmaceutical composition, or may be in separatepharmaceutical compositions. Each of the two or more pharmaceuticalagents may be administered through the same or different routes ofadministration. Co-administration encompasses administration in parallelor sequentially.

“Administered concomitantly” refers to the administration of two agentsat the same therapeutic time frame, in any manner in which thepharmacological effects of both are manifest in the patient at the sametime. Concomitant administration does not require that both agents beadministered in a single pharmaceutical composition, in the same dosageform, or by the same route of administration.

“Therapeutic lifestyle change” means dietary and lifestyle changesintended to lower cholesterol and reduce the risk of developing heartdisease, and includes recommendations for dietary intake of total dailycalories, total fat, saturated fat, polyunsaturated fat, monounsaturatedfat, carbohydrate, protein, cholesterol, insoluble fiber, as well asrecommendations for physical activity.

“Statin” means a pharmaceutical agent that inhibits the activity ofHMG-CoA reductase.

“HMG-CoA reductase inhibitor” means a pharmaceutical agent that actsthrough the inhibition of the enzyme HMG-CoA reductase.

“Cholesterol absorption inhibitor” means a pharmaceutical agent thatinhibits the absorption of exogenous cholesterol obtained from diet.

“LDL apheresis” means a form of apheresis by which LDL-C is removed fromblood. Typically, a individual's blood is removed from a vein, andseparated into red cells and plasma. LDL-C is filtered out of the plasmaprior to return of the plasma and red blood cells to the individual.

“MTP inhibitor” means a pharmaceutical agent that inhibits the enzymemicrosomal triglyceride transfer protein.

“Low density lipoprotein-cholesterol (LDL-C)” means cholesterol carriedin low density lipoprotein particles. Concentration of LDL-C in serum(or plasma) is typically quantified in mg/dL or nmol/L. “Serum LDL-C”and “plasma LDL-C” mean LDL-C in the serum and plasma, respectively.

“Very low density lipoprotein-cholesterol (VLDL-C)” means cholesterolassociated with very low density lipoprotein particles. Concentration ofVLDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L.“Serum VLDL-C” and “plasma VLDL-C” mean VLDL-C in the serum or plasma,respectively.

“Intermediate low density lipoprotein-cholesterol (IDL-C)” meanscholesterol associated with intermediate density lipoprotein.Concentration of IDL-C in serum (or plasma) is typically quantified inmg/dL or nmol/L. “Serum IDL-C” and “plasma IDL-C” mean IDL-C in theserum or plasma, respectively.

“Non-high density lipoprotein-cholesterol (Non-HDL-C)” means cholesterolassociated with lipoproteins other than high density lipoproteins, andincludes, without limitation, LDL-C, VLDL-C, and IDL-C.

“High density lipoprotein-C(HDL-C)” means cholesterol associated withhigh density lipoprotein particles. Concentration of HDL-C in serum (orplasma) is typically quantified in mg/dL or nmol/L. “Serum HDL-C” and“plasma HDL-C” mean HDL-C in the serum and plasma, respectively.

“Total cholesterol” means all types of cholesterol, including, but notlimited to, LDL-C, HDL-C, IDL-C and VLDL-C. Concentration of totalcholesterol in serum (or plasma) is typically quantified in mg/dL ornmol/L.

“Lipoprotein(a)” or “Lp(a)” means a lipoprotein particle that iscomprised of LDL-C, an apolipoprotein(a) particle, and anapolipoproteinB-100 particle.

“ApoA1” means apolipoprotein-A1 protein in serum. Concentration of ApoA1in serum is typically quantified in mg/dL or nmol/L.

“ApoB:ApoA1 ratio” means the ratio of ApoB concentration to ApoA1concentration.

“ApoB-containing lipoprotein” means any lipoprotein that hasapolipoprotein B as its protein component, and is understood to includeLDL, VLDL, IDL, and lipoprotein(a).

“Small LDL particle” means a subclass of LDL particles characterized bya smaller, denser size compared to other LDL particles. In certainembodiments, large LDL particles are 23-27 nm in diameter. In certainembodiments, intermediate LDL particles are 21.2-23 nm in diameter. Incertain embodiments, small LDL particles are 18-21.2 nm in diameter. Incertain embodiments, particle size is measured by nuclear magneticresonance analysis.

“Small VLDL particle” means a subclass of VLDL particles characterizedby a smaller, denser size compared to other VLDL particles. In certainembodiments, large VLDL particles are greater than 60 nm in diameter. Incertain embodiments, medium VLDL particles are 35-60 nm in diameter. Incertain embodiments, small VLDL particles are 27-35 nm in diameter. Incertain embodiments, particle size is measured by nuclear magneticresonance analysis.

“Triglycerides” means lipids that are the triesters of glycerol. “Serumtriglycerides” mean triglycerides present in serum. “Livertriglycerides” mean triglycerides present in liver tissue.

“Serum lipids” mean cholesterol and triglycerides in the serum.

“Elevated total cholesterol” means total cholesterol at a concentrationin an individual at which lipid-lowering therapy is recommended, andincludes, without limitation, elevated LDL-C″, “elevated VLDL-C,”“elevated IDL-C,” and “elevated non-HDL-C.” In certain embodiments,total cholesterol concentrations of less than 200 mg/dL, 200-239 mg/dL,and greater than 240 mg/dL are considered desirable, borderline high,and high, respectively. In certain embodiments, LDL-C concentrations of100 mg/dL, 100-129 mg/dL, 130-159 mg/dL, 160-189 mg/dL, and greater than190 mg/dL are considered optimal, near optimal/above optimal, borderlinehigh, high, and very high, respectively.

“Elevated triglyceride” means concentrations of triglyceride in theserum or liver at which lipid-lowering therapy is recommended, andincludes “elevated serum triglyceride” and “elevated livertriglyceride.” In certain embodiments, serum triglyceride concentrationof 150-199 mg/dL, 200-499 mg/dL, and greater than or equal to 500 mg/dLis considered borderline high, high, and very high, respectively.

“Elevated small LDL particles” means a concentration of small LDLparticles in an individual at which lipid-lowering therapy isrecommended.

“Elevated small VLDL particles” means a concentration of small VLDLparticles in an individual at which lipid-lowering therapy isrecommended.

“Elevated lipoprotein(a)” means a concentration of lipoprotein(a) in anindividual at which lipid-lowering therapy is recommended.

“Low HDL-C” means a concentration of HDL-C in an individual at whichlipid-lowering therapy is recommended. In certain embodimentslipid-lowering therapy is recommended when low HDL-C is accompanied byelevations in non-HDL-C and/or elevations in triglyceride. In certainembodiments, HDL-C concentrations of less than 40 mg/dL are consideredlow. In certain embodiments, HDL-C concentrations of less than 50 mg/dLare considered low.

“ApoB” means apolipoprotein B-100 protein. Concentration of ApoB inserum (or plasma) is typically quantified in mg/dL or nmol/L. “SerumApoB” and “plasma ApoB” mean ApoB in the serum and plasma, respectively.

“LDL/HDL ratio” means the ratio of LDL-C to HDL-C.

“Oxidized-LDL” or “Ox-LDL-C” means LDL-C that is oxidized followingexposure to free radicals.

“Individual having elevated LDL-C levels” means an individual who hasbeen identified by a medical professional (e.g., a physician) as havingLDL-C levels near or above the level at which therapeutic interventionis recommended, according to guidelines recognized by medicalprofessionals. Such an individual may also be considered “in need oftreatment” to decrease LDL-C levels.

“Individual having elevated apoB-100 levels” means an individual who hasbeen identified as having apoB-100 levels near or above the level atwhich therapeutic intervention is recommended, according to guidelinesrecognized by medical professionals. Such an individual may also beconsidered “in need of treatment” to decrease apoB-100 levels.

“Treatment of elevated LDL-C levels” means administration of anantisense compound targeted to a PCSK9 nucleic acid to an individualhaving elevated LDL-C levels.

“Treatment of atherosclerosis” means administration of an antisensecompound targeted to a PCSK9 nucleic acid to an individual who, basedupon a physician's assessment, has or is likely to have atherosclerosis.“Prevention of atherosclerosis” means administration of an antisensecompound targeted to a PCSK9 nucleic acid to an individual who, basedupon a physician's assessment, is susceptible to atherosclerosis.

“Ameliorate” means to make better or improve a detrimental condition inan individual.

“Target nucleic acid,” “target RNA,” “target RNA transcript” and“nucleic acid target” all mean any nucleic acid capable of beingtargeted by antisense compounds.

“PCSK9 nucleic acid” means any nucleic acid encoding PCSK9. For example,in certain embodiments, a PCSK9 nucleic acid includes, withoutlimitation, a DNA sequence encoding PCSK9, an RNA sequence transcribedfrom DNA encoding PCSK9, and an mRNA sequence encoding PCSK9. “PCSK9mRNA” means an mRNA encoding a PCSK9 protein.

“Targeting” means the process of design and selection of an antisensecompound that will specifically hybridize to a target nucleic acid andinduce a desired effect.

“Targeted” means having a nucleobase sequence that will allow specifichybridization of an antisense compound to a target nucleic acid toinduce a desired effect. In certain embodiments, a desired effect isreduction of a target nucleic acid. In certain such embodiments, adesired effect is reduction of PCSK9 mRNA.

“Antisense inhibition” means reduction of a target nucleic acid levelsin the presence of an antisense compound complementary to a targetnucleic acid compared to target nucleic acid levels in the absence ofthe antisense compound.

“Target region” means a fragment of a target nucleic acid to which oneor more antisense compounds is targeted.

“Target segment” means the sequence of nucleotides of a target nucleicacid to which an antisense compound is targeted. “5′ target site” refersto the 5′-most nucleotide of a target segment. “3′ target site” refersto the 3′-most nucleotide of a target segment.

“Active target region” means a target region to which one or more activeantisense compounds is targeted. “Active antisense compounds” meansantisense compounds that reduce target nucleic acid levels.

“Hybridization” means the annealing of complementary nucleic acidmolecules. In certain embodiments, complementary nucleic acid moleculesinclude, but are not limited to, an antisense compound and a nucleicacid target. In certain such embodiments, complementary nucleic acidmolecules include, but are not limited to, an antisense oligonucleotideand a nucleic acid target

“Stringent hybridization conditions” means conditions under which anucleic acid molecule, such as an antisense compound, will hybridize toa target nucleic acid sequence, but to a minimal number of othersequences.

“Specifically hybridizable” means an antisense compound that hybridizesto a target nucleic acid to induce a desired effect, while exhibitingminimal or no effects on non-target nucleic acids.

“Complementarity” means the capacity for pairing between nucleobases ofa first nucleic acid and a second nucleic acid.

“Fully complementary” means each nucleobase of a first nucleic acid iscapable of pairing with each nucleobase of a second nucleic acid. Incertain embodiments, a first nucleic acid is an antisense compound and atarget nucleic acid is a second nucleic acid. In certain suchembodiments, an antisense oligonucleotide is a first nucleic acid and atarget nucleic acid is a second nucleic acid.

“Non-complementary nucleobase” means a nucleobase of first nucleic acidthat is not capable of pairing with the corresponding nucleobase of atarget nucleic acid. “Mismatch” a nucleobase of first nucleic acid thatis not capable of pairing with the corresponding nucleobase of a targetnucleic acid.

“Portion” means a defined number of contiguous (i.e., linked)nucleobases. In certain embodiments, a portion is a defined number ofcontiguous nucleobases of a target nucleic acid. In certain embodiments,a portion is a defined number of contiguous nucleobases of an antisensecompound.

“Oligomeric compound” means a polymer of linked monomeric subunits whichis capable of hybridizing to at least a region of an RNA molecule.

“Antisense compound” means an oligomeric compound that is capable ofundergoing hybridization to a target nucleic acid through hydrogenbonding.

“Oligonucleotide” means a polymer of linked nucleosides each of whichcan be modified or unmodified, independent one from another.

“Oligonucleoside” means an oligonucleotide in which the internucleosidelinkages do not contain a phosphorus atom.

“Unmodified nucleotide” means a nucleotide composed of naturallyoccurring nucleobases, sugar moieties and internucleoside linkages. Incertain embodiments, an unmodified nucleotide is an RNA nucleotide(i.e., β-D-ribonucleosides) or a DNA nucleotide (i.e.,β-D-deoxyribonucleoside).

“Antisense oligonucleotide” means a single-stranded oligonucleotidehaving a nucleobase sequence that permits hybridization to acorresponding region of a target nucleic acid.

“Motif” means the pattern of unmodified and modified nucleosides in anantisense compound.

“Chimeric antisense compound” means an antisense compound that has atleast 2 chemically distinct regions, each region having a plurality ofsubunits.

A “gapmer” means an antisense compound in which an internal positionhaving a plurality of nucleotides that supports RNaseH cleavage ispositioned between external regions having one or more nucleotides thatare chemically distinct from the nucleosides of the internal region.

A “gap segment” means the plurality of nucleotides that make up theinternal region of a gapmer.

A “wing segment” means the external region of a gapmer.

“Gap-widened” means an antisense compound has a gap segment of 12 ormore contiguous 2′-deoxyribonucleotides positioned between 5′ and 3′wing segments having from one to six nucleotides having modified sugarmoieties.

“Nucleoside” means a nucleobase linked to a sugar.

“Nucleobase” means a heterocyclic base moiety capable of pairing with abase of another nucleic acid.

“Nucleotide” means a nucleoside having a phosphate group covalentlylinked to the sugar portion of the nucleoside.

“Nucleobase sequence” means the order of contiguous nucleobasesindependent of any sugar, linkage, and/or nucleobase modification.

“Modified nucleotide” means a nucleotide having, independently, amodified sugar moiety, modified internucleoside linkage, or modifiednucleobase.

“Modified nucleoside” means a nucleotide having, independently, amodified sugar moiety or modified nucleobase.

“Contiguous nucleobases” means nucleobases immediately adjacent to eachother.

“Modified oligonucleotide” means an oligonucleotide comprising amodified internucleoside linkage, a modified sugar, and/or a modifiednucleobase.

“Single-stranded modified oligonucleotide” means a modifiedoligonucleotide which is not hybridized to a complementary strand.

“Internucleoside linkage” refers to the chemical bond betweennucleosides.

“Linked nucleoside” means adjacent nucleosides which are bondedtogether.

“Linked deoxynucleoside” means a nucleic acid base (A, G, C, T, U)substituted by deoxyribose linked by a phosphate ester to form anucleotide.

“Naturally occurring internucleoside linkage” means a 3′ to 5′phosphodiester linkage.

“Modified internucleoside linkage” means substitution and/or any changefrom a naturally occurring internucleoside bond. In certain instances,the modified internucleoside linkage refers to a phosphodiesterinternucleoside bond.

“Phosphorothioate linkage” means a linkage between nucleosides where thephosphodiester bond is modified by replacing one of the non-bridgingoxygen atoms with a sulfur atom. A phosphorothioate linkage is amodified internucleoside linkage.

“Modified sugar moiety” means substitution and/or any change from anatural sugar moiety. For the purposes of this disclosure, a “naturalsugar moiety” is a sugar moiety found in DNA (2′-H) or RNA (2′-OH).

“Modified sugar” refers to a substitution and/or any change from anatural sugar.

“Bicyclic sugar” means a furosyl ring modified by the bridging of thetwo non-geminal ring atoms. A bicyclic sugar is a modified sugar.

“Modified nucleobase” means any nucleobase other than adenine, cytosine,guanine, thymidine, or uracil. An “unmodified nucleobase” means thepurine bases adenine (A) and guanine (G), and the pyrimidine basesthymine (T), cytosine (C) and uracil (U).

“Modified sugar moiety” means a sugar moiety having a substitutionand/or any change from a natural sugar moiety.

“Natural sugar moiety” means a sugar moiety found in DNA (2′-H) or RNA(2′-OH).

“2′-O-methoxyethyl sugar moiety” means a 2′-substituted furosyl ringhaving a 2′-O(CH2)₂—OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituentgroup.

“2′-O-methoxyethyl nucleotide” means a nucleotide comprising a2′-O-methoxyethyl modified sugar moiety.

“2′-O-methoxyethyl” refers to an O-methoxy-ethyl modification of the 2′position of a furosyl ring. A 2′-O-methoxyethyl modified sugar is amodified sugar.

“Bicyclic nucleic acid sugar moiety” means a furosyl ring modified bythe bridging of two non-geminal ring atoms.

“5-methylcytosine” means a cytosine modified with a methyl groupattached to the 5′ position. A 5-methylcytosine is a modifiednucleobase.

“Prodrug” means a therapeutic agent that is prepared in an inactive formthat is converted to an active form (i.e., drug) within the body orcells thereof by the action of endogenous enzymes or other chemicalsand/or conditions.

“Pharmaceutically acceptable salts” means physiologically andpharmaceutically acceptable salts of antisense compounds, i.e., saltsthat retain the desired biological activity of the parentoligonucleotide and do not impart undesired toxicological effectsthereto.

“Salts” refers to physiologically and pharmaceutically acceptable saltsof antisense compounds, i.e., salts that retain the desired biologicalactivity of the parent oligonucleotide and do not impart undesiredtoxicological effects thereto.

“Cures” means a method or course that restores health or a prescribedtreatment for an illness.

“Slows progression” means decrease in the development of the saiddisease.

“Cap structure” or “terminal cap moiety” means chemical modifications,which have been incorporated at either terminus of an antisensecompound.

“ISIS 301012” means a lipid-lowering agent that is an antisenseoligonucleotide having the sequence “GCCTCAGTCTGCTTCGCACC” (SEQ ID NO:457), where each internucleoside linkage is a phosphorothioateinternucleoside linkage, each cytosine is a 5-methylcytosine,nucleotides 6-15 are 2′-deoxynucleotides, and nucleotides 1-5 and 16-20are 2′-O-methoxyethyl nucleotides. ISIS 301012 does not target PCSK9. Itis used herein as a non-PCSK9 control.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts the location of PCSK9 mRNA target sequences of exemplaryantisense compounds.

FIG. 2 depicts the results of a five-point dose response experiment withexemplary antisense compounds in HEK-293 cells. The antisense compoundsare identified by Isis number. The bars from left to right representPCSK9 expression in relative units at 0 nM, 25 nM, 50 nM, 100 nM, 200nM, and 300 nM antisense oligonucleotides.

FIG. 3 depicts the results of a five-point dose response experiment withexemplary antisense compounds in cyno primary hepatocytes. The antisensecompounds are identified by Isis number. The bars from left to rightrepresent PCSK9 expression in relative units at 300 nM, 150 nM, 50 nM,25 nM, and 10 nM antisense oligonucleotides. Control values are shown inthe rightmost two bars.

FIG. 4 depicts a reduction of murine PCSK9 mRNA expression (relativeunits) in the liver of h-apoB/CETP transgenic mice when administeredIsis 394816. Isis 394816 was administered by intraperitoneal injectionfor 6 weeks at a dosing regimen of 20 mg/kg/wk or 50 mg/kg/wk. Isis329993 was administered at 50 mg/kg/wk as a control.

FIG. 5, Panel A, depicts inhibition of plasma LDL-C (mg/dL) in miceadministered Isis 394816 at 20 mg/kg/wk or 50 mg/kg/wk. FIG. 5, Panel B,depicts inhibition of liver TG (mg/gram) in mice administered Isis394816 at 20 mg/kg/wk or 50 mg/kg/wk.

FIG. 6 depicts the dose-dependent inhibition (relative units) of liverPCSK9 mRNA, plasma LDL-C, and liver TG in response to the indicateddoses of Isis 394816.

FIG. 7 depicts a correlation between LDL-C lowering and PCSK9 mRNAinhibition, using the data from FIG. 6.

FIG. 8 depicts the accumulation of exemplary antisense compounds in rattissues (μg/g tissue) following administration of antisense compounds.The antisense compounds are identified by Isis number. The bars fromleft to right represent antisense compound accumulation in kidney andliver.

FIG. 9 depicts the accumulation of the identified antisense compounds intissues (μg/g tissue) following administration to a cyno monkey.Accumulation at each dosing concentration (15 mg/kg and 30 mg/kg) isshown in cyno kidney (left side bar) and cyno liver (right side bar).

FIG. 10 depicts PCSK9 mRNA levels (relative units) followingadministration of the identified antisense compounds to a cyno monkey.Results are shown at each dosing concentration (15 mg/kg and 30 mg/kg).

FIG. 11 depicts hepatic PCSK9 mRNA levels (relative units) detected byNorthern analysis. Cyno monkeys were administered Isis 405879(“BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg.

FIG. 12 depicts plasma LDL-C levels (mg/dL) following administration ofIsis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg.Left side bars represent baseline levels of LDL-C in control animals;right side bars represent LDL-C following administration of antisensecompounds.

FIG. 13 depicts the ratio of plasma HDL/LDL following administration ofIsis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg.Left side bars represent baseline HDL/LDL in control animals; right sidebars represent HDL/LDL following administration of antisense compounds.

FIG. 14 depicts the effect of BMS-844421 and control ASOs on liver humanPCSK9 mRNA in transgenic mice (6 wk).

FIG. 15 depicts the effect of BMS-844421 and control ASOs on plasmalevels of human PCSK9 protein in transgenic mice (6 wk).

FIG. 16 depicts plasma hPCSK9 at baseline (FIG. 16A), 3 wk (FIG. 16B),and 6 wk (FIG. 16C) treatment.

FIG. 17 depicts the BMS-844421 dose response for plasma hPCSK9 levels intransgenic mice at 6 weeks (wk).

FIG. 18 depicts the BMS-844421 dose response for liver mRNA and plasmaprotein (hPCSK9).

FIG. 19 depicts the dose response for plasma hPCSK9 in male and femaletransgenic mice.

FIG. 20 depicts the dose response for liver hPCSK9 mRNA in male versusfemale transgenic mice.

FIG. 21 depicts the effect of the ASOs on liver endogenous PCSK9 mRNA inthe transgenic mouse after 6 weeks of administration. The numbersfollowing the hyphens provide the mg ASO/kg body weight dose of eachASO. For example, “844421-30” is the data for BMS-84421 administered totest animals at 30 mg/kg.

FIG. 22 depicts the effect of ASOs on liver endogenous HMGCoA reductasemRNA in the transgenic mouse after 6 weeks of administration.

FIG. 23 depicts LDLR protein concentration following a 6 wk ASOtreatment, as assayed by a Western blot.

FIG. 24 depicts the quantification of the Western blot shown in FIG. 23.The LDLR band is normalized to a transferring receptor (TR) band, asobserved using anti-LDLR and anti-TR antibodies, respectively.

FIG. 25 depicts the Liver PCSK9 mRNA concentration in cyno monkeys after13 weeks of treatment. The numbers in parentheses are administered dosesin mg/kg.

FIG. 26 depicts the concentration of BMS-844421 in the liver of cynomonkeys after 13 weeks of treatment.

FIG. 27 depicts the average serum LDL-C levels in the cyno monkey study.The numbers in parentheses are administered doses in mg/kg.

FIG. 28A depicts the pharmacodynamic responses in serum lipid assays forthe cyno 2.5 and 5 wk ASO studies, as displayed in a scatterplot ofindividual animal data. FIG. 28B depicts the average ratio of HDL/LDL inBMS-844421-treated cynos at 2.5 and 5 wk. The numbers in parentheses inFIG. 28B are administered doses in mg/kg.

FIG. 29 depicts downregulation of PCSK9 target in cyno monkeys at 13 wkin the toxicology study of liver mRNA and plasma PCSK9.

DETAILED DESCRIPTION Overview

Elevated levels of LDL-cholesterol (HDL-C) are recognized as a majorindependent risk factor for coronary heart disease (CHD). Even inindividuals undergoing aggressive treatment with currently availablecholesterol-lowering agents to reduce LDL-cholesterol (LDL-C) levels,coronary events still occur, and elevated LDL-C levels remain a majorrisk factor for coronary heart disease in these individuals.Furthermore, many individuals undergoing LDL-lowering therapy do notreach their target LDL-C levels, and thus remain at risk for CHD.Accordingly, there is a need for additional LDL-C lowering agents.

The antisense compounds and methods provided herein are useful for thetreatment of hypercholesterolemia. Treatment of hypercholesterolemiaencompasses a therapeutic regimen that results in a clinically desirableoutcome. For example, the antisense compounds and methods providedherein are useful for the treatment of elevated cholesterol, such aselevated LDL-C. In addition, the antisense compounds and methodsprovided herein may be used to reduce the risk of CHD, in individualsexhibiting one or more risk factors for CHD. Furthermore, the antisensecompounds and methods provided herein may be used to treat and/orprevent atherosclerosis.

As illustrated herein, administration of an antisense oligonucleotidetargeted to PCSK9 to animals fed a high-fat diet (an experimental modelof hyperlipidemia) resulted in antisense inhibition of PCSK9,upregulation of the LDL-R, reduction of LDL-C levels, and reduction ofliver triglycerides. Thus, it is demonstrated that in an experimentalmodel of hyperlipidemia, antisense inhibition of PCSK9 results inlowering LDL-C levels. Accordingly, provided herein are methods for thetreatment of reducing LDL-C levels through the administration of anantisense compound targeted to a PCSK9 nucleic acid. Increased LDL-Clevels are considered a risk factor for CHD, and are also linked toatherosclerosis. Accordingly, also provided herein are methods for thereduction of CHD risk, and for the prevention and/or treatment ofatherosclerosis. Also provided herein are methods for the treatment ofconditions characterized by elevated liver triglycerides, such ashepatic steatosis.

In a particular embodiment, methods comprise the use of antisensecompounds targeted to particularly advantageous sequences within a PCSK9nucleic acid. The PCSK9 target sequences are selected on the basis ofsuperior results shown in one or more selection criteria. The presentlydisclosed antisense compounds all exhibit high in vitro efficacy,determined using a battery of cell models to measure antisenseregulation of PCSK9 expression. Further, preferred antisense compoundsexhibit relatively high in vivo efficacy, determined using a transgenicmouse model that expresses human PCSK9 or a cynomolgus (“cyno”) monkeymodel. The antisense compound ISIS 405879 (SEQ ID NO: 248) displayssuperior results in suppressing PCSK9 mRNA expression in the liver of ahost individual.

The present antisense compounds are tolerated to different extents whenadministered to a host individual. Increased tolerability can depend ona number of factors, including, but not limited to, the nucleotidesequence of the antisense compound, chemical modifications to thenucleotides, the particular motif of unmodified and modified nucleosidesin the antisense compound, or combinations thereof. Antisense compoundsthat exhibit increased or superior tolerability are particularlypreferred in the present methods. The present antisense compounds alsocan show different pharmacokinetic properties, depending on theirnucleotide sequence, chemical modifications, motifs, or combinationsthereof. Particularly preferred antisense compounds also exhibitfavorable pharmacokinetics when administered to a host individual.

Certain Indications

In certain embodiments, the invention provides methods of treating anindividual comprising administering one or more pharmaceuticalcompositions of the present invention. In certain embodiments, theindividual has hypercholesterolemia, mixed dyslipidemia,atherosclerosis, a risk of developing atherosclerosis, coronary heartdisease, a history of coronary heart disease, early onset coronary heartdisease, acute coronary syndrome, one or more risk factors for coronaryheart disease, type II diabetes, type II diabetes with dyslipidemia,dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia,hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fattyliver disease.

Guidelines for lipid-lowering therapy were established in 2001 by AdultTreatment Panel III (ATP III) of the National Cholesterol EducationProgram (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004,110, 227-239). The guidelines include obtaining a complete lipoproteinprofile, typically after a 9 to 12 hour fast, for determination ofLDL-C, total cholesterol, and HDL-C levels. According to the mostrecently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189mg/dL, and greater than or equal to 190 mg/dL are considered borderlinehigh, high, and very high, respectively. Total cholesterol levels of200-239 and greater than or equal to 240 mg/dL are considered borderlinehigh and high, respectively. HDL-C levels of less than 40 mg/dL areconsidered low.

In certain embodiments, the individual has been identified as in need oflipid-lowering therapy. In certain such embodiments, the individual hasbeen identified as in need of lipid-lowering therapy according to theguidelines established in 2001 by Adult Treatment Panel III (ATP III) ofthe National Cholesterol Education Program (NCEP), and updated in 2004(Grundy et al., Circulation, 2004, 110, 227-239). In certain suchembodiments, the individual in need of lipid-lowering therapy has LDL-Cabove 190 mg/dL. In certain such embodiments, the individual in need oflipid-lowering therapy has LDL-C above 160 mg/dL. In certain suchembodiments, the individual in need of lipid-lowering therapy has LDL-Cabove 130 mg/dL. In certain such embodiments, the individual in need oflipid-lowering therapy has LDL-C above 100 mg/dL. In certain suchembodiments, the individual in need of lipid-lowering therapy shouldmaintain LDL-C below 160 mg/dL. In certain such embodiments, theindividual in need of lipid-lowering therapy should maintain LDL-C below130 mg/dL. In certain such embodiments, the individual in need oflipid-lowering therapy should maintain LDL-C below 100 mg/dL. In certainsuch embodiments, the individual should maintain LDL-C below 70 mg/dL oreven below 50 mg/dL.

In certain embodiments, the invention provides methods for reducing ApoBin an individual. In certain embodiments, the invention provides methodsfor reducing ApoB-containing lipoprotein in an individual. In certainembodiments, the invention provides methods for reducing LDL-C in anindividual. In certain embodiments, the invention provides methods forreducing VLDL-C in an individual. In certain embodiments, the inventionprovides methods for reducing IDL-C in an individual. In certainembodiments, the invention provides methods for reducing non-HDL-C in anindividual. In certain embodiments the invention provides methods forreducing Lp(a) in an individual. In certain embodiments, the inventionprovides methods for reducing serum triglyceride in an individual. Incertain embodiments, the invention provides methods for reducing livertriglyceride in an individual. In certain embodiments, the inventionprovides methods for reducing Ox-LDL-C in an individual. In certainembodiments, the invention provides methods for reducing small LDLparticles in an individual. In certain embodiments, the inventionprovides methods for reducing small VLDL particles in an individual. Incertain embodiments, the invention provides methods for reducingphospholipids in an individual. In certain embodiments, the inventionprovides methods for reducing oxidized phospholipids in an individual.

In certain embodiments, the methods provided by the present invention donot lower HDL-C. In certain embodiments, the methods provided by thepresent invention do not result in accumulation of lipids in the liver.

In one embodiment are methods for decreasing LDL-C levels, oralternatively methods for treating hypercholesterolemia, byadministering to an individual suffering from elevated LDL-C levels atherapeutically effective amount of an antisense compound targeted to aPCSK9 nucleic acid. In another embodiment, a method of decreasing LDL-Clevels comprises selecting an individual in need of a decrease in LDL-Clevels, and administering to the individual a therapeutically effectiveamount of an antisense compound targeted to a PCSK9 nucleic acid. In afurther embodiment, a method of reducing coronary heart disease riskincludes selecting an individual having elevated LDL-C levels and one ormore additional indicators of coronary heart disease risk, andadministering to the individual a therapeutically effective amount of anantisense compound targeted to a PCSK9 nucleic acid.

In other embodiments, the LDL-C level is from 100-129 mg/dL, from 130 to159 mg/dL, from 160-189 mg/dL, or greater than or equal to 190 mg/dL.

In one embodiment, administration of a therapeutically effective amountof an antisense compound targeted to a PCSK9 nucleic acid is accompaniedby monitoring of LDL-C levels in the serum of an individual, todetermine an individual's response to administration of the antisensecompound. An individual's response to administration of the antisensecompound is used by a physician to determine the amount and duration oftherapeutic intervention.

In one embodiment, administration of an antisense compound targeted to aPCSK9 nucleic acid results in LDL-C levels below 190 mg/dL, below 160mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50mg/dL. In another embodiment, administration of an antisense compoundtargeted to a PCSK9 nucleic acid decreases LDL-C by at least 15%, by atleast 25%, by at least 50%, by at least 60%, by at least 70%, by atleast 75%, by at least 80%, by at least 85%, by at least 90%, or by atleast 95%.

An individual having elevated LDL-C levels may also exhibit reducedHDL-C levels and/or elevated total cholesterol levels. Accordingly, inone embodiment a therapeutically effective amount of an antisensecompound targeted to a PCSK9 nucleic acid is administered to anindividual having elevated LDL-C levels, who also has reduced HDL-Clevels and/or elevated total cholesterol levels.

Individuals having elevated LDL-C levels may also exhibit elevatedtriglyceride levels. Accordingly, in one embodiment a therapeuticallyeffective amount of an antisense compound targeted to a PCSK9 nucleicacid is administered to an individual having elevated LDL-C levels, andalso having elevated triglyceride levels.

Atherosclerosis can lead to coronary heart disease, stroke, orperipheral vascular disease. Elevated LDL-C levels are considered a riskfactor in the development and progression of atherosclerosis.Accordingly, in one embodiment, a therapeutically effective amount of anantisense compound targeted to a PCSK9 nucleic acid is administered toan individual having atherosclerosis. In a further embodiment, atherapeutically effective amount of antisense compound targeted to aPCSK9 nucleic acid is administered to an individual susceptible toatherosclerosis. Atherosclerosis is assessed directly through routineimaging techniques such as, for example, ultrasound imaging techniquesthat reveal carotid intimomedial thickness. Accordingly, treatmentand/or prevention of atherosclerosis further include monitoringatherosclerosis through routine imaging techniques. In one embodiment,administration of an antisense compound targeted to a PCSK9 nucleic acidleads to a lessening of the severity of atherosclerosis, as indicatedby, for example, a reduction of carotid intimomedial thickness inarteries.

Measurements of cholesterol, lipoproteins and triglycerides are obtainedusing serum or plasma collected from an individual. Methods of obtainingserum or plasma samples are routine, as are methods of preparation ofthe serum samples for analysis of cholesterol, triglycerides, and otherserum markers.

A physician may determine the need for therapeutic intervention forindividuals in cases where more or less aggressive LDL-lowering therapyis needed. The practice of the methods herein may be applied to anyaltered guidelines provided by the NCEP, or other entities thatestablish guidelines for physicians used in treating any of the diseasesor conditions listed herein, for determining coronary heart disease riskand diagnosing metabolic syndrome.

In one embodiment, administration of an antisense compound targeted to aPCSK9 nucleic acid is parenteral administration. Parenteraladministration may be intravenous or subcutaneous administration.Accordingly, in another embodiment, administration of an antisensecompound targeted to a PCSK9 nucleic acid is intravenous or subcutaneousadministration. Administration may include multiple doses of anantisense compound targeted to a PCSK9 nucleic acid.

In certain embodiments, a pharmaceutical composition comprising anantisense compound targeted to PCSK9 is for use in therapy. In certainembodiments, the therapy is the reduction of LDL-C, ApoB, VLDL-C, IDL-C,non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C,small LDL particles, small VLDL, phospholipids, or oxidizedphospholipids in an individual. In certain embodiments, the therapy isthe treatment of hypercholesterolemia, mixed dyslipidemia,atherosclerosis, a risk of developing atherosclerosis, coronary heartdisease, acute coronary syndrome, a history of coronary heart disease,early onset coronary heart disease, one or more risk factors forcoronary heart disease, type II diabetes, type II diabetes withdyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia,hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, ornon-alcoholic fatty liver disease. In additional embodiments, thetherapy is the reduction of CHD risk. In certain aspects, the therapy isprevention of atherosclerosis. In certain embodiments, the therapy isthe prevention of coronary heart disease.

In certain embodiments, a pharmaceutical composition comprising anantisense compound targeted to PCSK9 is used for the preparation of amedicament for reducing LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a),serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles,small VLDL, phospholipids, or oxidized phospholipids in an individual.In certain embodiments pharmaceutical composition comprising anantisense compound targeted to PCKS9 is used for the preparation of amedicament for reducing coronary heart disease risk. In certainembodiments, an antisense compound targeted to PCSK9 is used for thepreparation of a medicament for the treatment of hypercholesterolemia,mixed dyslipidemia, atherosclerosis, a risk of developingatherosclerosis, coronary heart disease, a history of coronary heartdisease, early onset coronary heart disease, one or more risk factorsfor coronary heart disease, type II diabetes, type II diabetes withdyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia,hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, ornon-alcoholic fatty liver disease.

Certain Combination Therapies

In certain embodiments, one or more pharmaceutical compositions of thepresent invention are co-administered with one or more otherpharmaceutical agents. In certain embodiments, such one or more otherpharmaceutical agents are designed to treat the same disease orcondition as the one or more pharmaceutical compositions of the presentinvention. In certain embodiments, such one or more other pharmaceuticalagents are designed to treat a different disease or condition as the oneor more pharmaceutical compositions of the present invention. In certainembodiments, such one or more other pharmaceutical agents are designedto treat an undesired effect of one or more pharmaceutical compositionsof the present invention. In certain embodiments, one or morepharmaceutical compositions of the present invention are co-administeredwith another pharmaceutical agent to treat an undesired effect of thatother pharmaceutical agent. In certain embodiments, one or morepharmaceutical compositions of the present invention and one or moreother pharmaceutical agents are administered at the same time. Incertain embodiments, one or more pharmaceutical compositions of thepresent invention and one or more other pharmaceutical agents areadministered at different times. In certain embodiments, one or morepharmaceutical compositions of the present invention and one or moreother pharmaceutical agents are prepared together in a singleformulation. In certain embodiments, one or more pharmaceuticalcompositions of the present invention and one or more otherpharmaceutical agents are prepared separately. For example, acomposition may comprise a pharmaceutical agent for separate,sequential, or simultaneous administration with an antisense compound.

In certain embodiments, pharmaceutical agents that may beco-administered with a pharmaceutical composition of the presentinvention include lipid-lowering agents or LXR agonists. In certain suchembodiments, pharmaceutical agents that may be co-administered with apharmaceutical composition of the present invention include, but are notlimited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. Incertain such embodiments, the lipid-lowering agent is administered priorto administration of a pharmaceutical composition of the presentinvention. In certain such embodiments, the lipid-lowering agent isadministered following administration of a pharmaceutical composition ofthe present invention. In certain such embodiments the lipid-loweringagent is administered at the same time as a pharmaceutical compositionof the present invention. In certain such embodiments the dose of aco-administered lipid-lowering agent is the same as the dose that wouldbe administered if the lipid-lowering agent was administered alone. Incertain such embodiments the dose of a co-administered lipid-loweringagent is lower than the dose that would be administered if thelipid-lowering agent was administered alone. In certain such embodimentsthe dose of a co-administered lipid-lowering agent is greater than thedose that would be administered if the lipid-lowering agent wasadministered alone.

In certain embodiments, a co-administered lipid-lowering agent is aHMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoAreductase inhibitor is a statin. In certain such embodiments, the statinis selected from, for example, atorvastatin, simvastatin, pravastatin,fluvastatin, and rosuvastatin.

In certain embodiments, a co-administered lipid-lowering agent is acholesterol absorption inhibitor. In certain such embodiments,cholesterol absorption inhibitor is ezetimibe.

In certain embodiments, a co-administered lipid-lowering agent is aco-formulated HMG-CoA reductase inhibitor and cholesterol absorptioninhibitor. In certain such embodiments the co-formulated lipid-loweringagent is ezetimibe/simvastatin.

In certain embodiments, a co-administered lipid-lowering agent is amicrosomal triglyceride transfer protein inhibitor (MTP inhibitor).

In certain embodiments, a co-administered lipid-lowering agent is anoligonucleotide targeted to ApoB.

In certain embodiments, a co-administered pharmaceutical agent is a bileacid sequestrant. In certain such embodiments, the bile acid sequestrantis selected from cholestyramine, colestipol, and colesevelam.

In certain embodiments, a co-administered pharmaceutical agent is anicotinic acid. In certain such embodiments, the nicotinic acid isselected from immediate release nicotinic acid, extended releasenicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is afibric acid. In certain such embodiments, a fibric acid is selected fromgemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.

Further examples of pharmaceutical agents that may be co-administeredwith a pharmaceutical composition of the present invention include, butare not limited to, corticosteroids, including but not limited toprednisone; LXR agonists; immunoglobulins, including, but not limited tointravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen);anti-inflammatory agents, including, but not limited to non-steroidalanti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2,inhibitors); salicylates; antibiotics; antivirals; antifungal agents;antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins,sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics;hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin,progestin, somatostan, and thyroid hormones); immunomodulators; musclerelaxants; antihistamines; osteoporosis agents (e.g., biphosphonates,calcitonin, and estrogens); prostaglandins, antineoplastic agents;psychotherapeutic agents; sedatives; poison oak or poison sumacproducts; antibodies; and vaccines.

In certain embodiments, the pharmaceutical compositions of the presentinvention may be administered in conjunction with a lipid-loweringtherapy. In certain such embodiments, a lipid-lowering therapy istherapeutic lifestyle change. In certain such embodiments, alipid-lowering therapy is LDL apheresis.

Antisense Compounds

Oligomeric compounds include, but are not limited to, oligonucleotides,oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics,antisense compounds, antisense oligonucleotides, and siRNAs. Anoligomeric compound may be “antisense” to a target nucleic acid, meaningthat is capable of undergoing hybridization to a target nucleic acidthrough hydrogen bonding.

In certain embodiments, an antisense compound has a nucleobase sequencethat, when written in the 5′ to 3′ direction, comprises the reversecomplement of the target segment of a target nucleic acid to which it istargeted. In certain such embodiments, an antisense oligonucleotide hasa nucleobase sequence that, when written in the 5′ to 3′ direction,comprises the reverse complement of the target segment of a targetnucleic acid to which it is targeted.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid is 8 to 80, 12 to 50, 12 to 30 or 15 to 30 subunits inlength. In other words, antisense compounds are from 8 to 80, 12 to 50,12 to 30 or 15 to 30 linked subunits. In certain such embodiments, theantisense compounds are 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, or 30 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9nucleic acid is 12 to 30 nucleotides in length. In certain suchembodiments, an antisense oligonucleotide targeted to a PCSK9 nucleicacid is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides in length.

In certain embodiment, an antisense compound targeted to a PCSK9 nucleicacid is 15 to 30 subunits in length. In other words, antisense compoundsare from 15 to 30 linked subunits. In certain such embodiments, theantisense compounds are 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, or 30 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9nucleic acid is 15 to 30 nucleotides in length. In certain suchembodiments, an antisense oligonucleotide targeted to a PCSK9 nucleicacid is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or30 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid is 18 to 24 subunits in length. In other words, antisensecompounds are from 18 to 24 linked subunits. In one embodiment, theantisense compounds are 18, 19, 20, 21, 22, 23, or 24 subunits inlength.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9nucleic acid is 18 to 24 nucleotides in length. In certain suchembodiments, an antisense oligonucleotide targeted to a PCSK9 nucleicacid is 18, 19, 20, 21, 22, 23, or 24 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid is 19 to 22 subunits in length. In other words, antisensecompounds are from 19 to 22 linked subunits. This embodies antisensecompounds of 19, 20, 21, or 22 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9nucleic acid is 19 to 22 nucleotides in length. In certain suchembodiments, an antisense oligonucleotide targeted to a PCSK9 nucleicacid is 19, 20, 21, or 22 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid is 20 subunits in length. In certain such embodiments,antisense compounds are 20 linked subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9nucleic acid is 20 nucleotides in length. In certain such embodiments,an antisense oligonucleotide targeted to an PCSK9 nucleic acid is 20linked nucleotides in length.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence target a regionidentified herein.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid may target the following nucleotide regions of SEQ ID NO:1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581,591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628,602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743,782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909,860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173,960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004,985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026,1000-1027, 1001-1028, 1002-1029, 1003-1029, 1004-1029, 1005-1029,1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115,1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444,1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377,1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602,1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624,1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591,1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706,1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765,1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878,1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943,1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948,1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952,1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206,2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334,2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625,2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927,2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526,410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622,600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647,638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917,835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909,923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064,970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024,997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021,1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034,1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251,1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345,1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416,1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519,1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568,1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592,1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666,1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876,1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879,1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945,1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950,1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059,2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332,2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334,2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855,2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013,3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid may target the following nucleotide regions of SEQ ID NO:2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579,2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618,5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623,6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565,6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602,6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584,8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233,9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491,12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847,13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423,14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650,14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915,14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963,14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290,15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321,15294-15321, 15295-15322, 15296-15323, 15297-15323, 15298-15323,15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490,15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286,18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668,18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080,19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009,19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070,20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804,20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697,20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736,20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748,20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014,21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152,21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608,21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311,22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163,22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227,22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231,22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236,22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311,23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926,25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303,26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141,26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161,26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662,26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or27350-27376.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575,2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638,3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463,6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562,6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567,6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621,6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967,9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235,10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947,13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141,14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513,14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700,14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943,14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998,15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318,15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322,15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323,15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490,16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580,18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730,18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981,19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042,20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207,20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781,20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781,20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745,20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752,20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152,21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211,21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227,22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163,22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163,22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228,22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233,22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239,22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134,24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013,26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141,26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141,26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335,26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736,26790-26820, 27034-27263, 27279-27303, or 27350-27376.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid may target the following nucleotide regions of SEQ ID NO:3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897,915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302,1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429,1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516,1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554,1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1558,1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602,1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781,1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198,1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127,1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057,1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019,1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025,2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131,2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381,2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589,2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589,2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174,3083-3110, 3155-3184, 3238-3268, 3482-3711, 3727-3751, or 3798-3824.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419,451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124,1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447,1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435,1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541,1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556,1530-1557, 1531-1557, 1532-1558, 1533-1559, 1534-1559, 1535-1559,1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645,1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781,1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867,1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971,1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999,1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024,1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139,2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177,2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587,2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589,2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880,2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268,3482-3711, 3727-3751, or 3798-3824.

In certain embodiments, an antisense compound targeted to a PCSK9nucleic acid may target the following nucleotide regions of SEQ ID NO:1: 320-405, 441-445, 527-544, 582-590, 744-781, 811-820, 918-922,953-959, 1034-1036, 1152-1153, 1174-1199, 1251-1272, 1445-1464,1603-1604, 1625-1627, 1707-1734, 1766-1811, 1832-1848, 1880-1904,1956-1961, 1982-1939, 2030-2039, 2060-2099, 2140-2149, 2170-2186,2207-2304, 2355-2409, 2435-2503, 2529-2581, 2626-2648, 2669-2749,2770-2827, 2856-2876, 2891-2899, 2930-2982, 3014-3226, 3253-3436,3457-3471, or 3497-3542.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 1: 320-405, 441-445, 527-544,582-590, 744-781, 811-820, 918-922, 953-959, 1034-1036, 1152-1153,1174-1199, 1251-1272, 1445-1464, 1603-1604, 1625-1627, 1707-1734,1766-1811, 1832-1848, 1880-1904, 1956-1961, 1982-1939, 2030-2039,2060-2099, 2140-2149, 2170-2186, 2207-2304, 2355-2409, 2435-2503,2529-2581, 2626-2648, 2669-2749, 2770-2827, 2856-2876, 2891-2899,2930-2982, 3014-3226, 3253-3436, 3457-3471, or 3497-3542.

In certain embodiments, a shortened or truncated antisense compoundtargeted to a PCSK9 nucleic acid has a single subunit deleted from the5′ end (5′ truncation), or alternatively from the 3′ end (3′truncation). A shortened or truncated antisense compound targeted to aPCSK9 nucleic acid may have two subunits deleted from the 5′ end, oralternatively may have two subunits deleted from the 3′ end, of theantisense compound. Alternatively, the deleted nucleosides may bedispersed throughout the antisense compound, for example, in anantisense compound having one nucleoside deleted from the 5′ end and onenucleoside deleted from the 3′ end.

When a single additional subunit is present in a lengthened antisensecompound, the additional subunit may be located at the 5′ or 3′ end ofthe antisense compound. When two are more additional subunits arepresent, the added subunits may be adjacent to each other, for example,in an antisense compound having two subunits added to the 5′ end (5′addition), or alternatively to the 3′ end (3′ addition), of theantisense compound. Alternatively, the added subunits may be dispersedthroughout the antisense compound, for example, in an antisense compoundhaving one subunit added to the 5′ end and one subunit added to the 3′end.

It is possible to increase or decrease the length of an antisensecompound, such as an antisense oligonucleotide, and/or introducemismatch bases without eliminating activity. For example, in Woolf etal. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series ofantisense oligonucleotides 13-25 nucleobases in length were tested fortheir ability to induce cleavage of a target RNA in an oocyte injectionmodel. Antisense oligonucleotides 25 nucleobases in length with 8 or 11mismatch bases near the ends of the antisense oligonucleotides were ableto direct specific cleavage of the target mRNA, albeit to a lesserextent than the antisense oligonucleotides that contained no mismatches.Similarly, target specific cleavage was achieved using 13 nucleobaseantisense oligonucleotides, including those with 1 or 3 mismatches.

Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001)demonstrated the ability of an oligonucleotide having 100%complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xLmRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and invivo. Furthermore, this oligonucleotide demonstrated potent anti-tumoractivity in vivo.

Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a seriesof tandem 14 nucleobase antisense oligonucleotides, and a 28 and 42nucleobase antisense oligonucleotides comprised of the sequence of twoor three of the tandem antisense oligonucleotides, respectively, fortheir ability to arrest translation of human DHFR in a rabbitreticulocyte assay. Each of the three 14 nucleobase antisenseoligonucleotides alone was able to inhibit translation, albeit at a moremodest level than the 28 or 42 nucleobase antisense oligonucleotides.

PCT/US2007/068404 describes incorporation of chemically-modifiedhigh-affinity nucleotides into short antisense compounds about 8-16nucleobases in length and that such compounds are useful in thereduction of target RNAs in animals with increased potency and improvedtherapeutic index.

In certain embodiments, antisense compounds targeted to a PCSK9 nucleicacid are short antisense compounds. In certain embodiments, such shortantisense compounds are oligonucleotide compounds. In certainembodiments, such short antisense compounds are about 8 to 16,preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14nucleotides in length and comprises a gap region flanked on each side bya wing, wherein each wing independently consists of 1 to 3 nucleotides.Preferred motifs include but are not limited to wing —deoxy gap—wingmotifs selected from 3-10-3, 2-10-3, 2-10-2, 1-10-1, 2-8-2, 1-8-1, 3-6-3or 1-6-1.

Antisense compounds targeted to a PCSK9 nucleic acid are synthesized invitro and do not include antisense compositions of biological origin, orgenetic vector constructs designed to direct the in vivo synthesis ofantisense molecules.

In certain embodiments, an antisense compound is targeted to a region ofa PCSK9 nucleic acid that does not contain a single nucleotidepolymorphism (SNPs). In certain embodiments, an antisense compound istargeted to a region of a PCSK9 nucleic acid that does contain a singlenucleotide polymorph (SNPs). A single nucleotide polymorphism refers topolymorphisms that are the result of a single nucleotide alteration orthe existence of two or more alternative sequences which can be, forexample, different allelic forms of a gene. A polymorphism may compriseone or more base changes including, for example, an insertion, a repeat,or a deletion. In certain embodiments, an antisense oligonucleotidetargeted to a PCSK9 nucleic acid overlaps with a SNP at the followingpositions: 428, 432, 449, 996, 1011, 1044, 1317, 1565, 1617, 1618, 1671,1711, 1722, 1836, 1911. In certain embodiments, the compounds providedherein that target a region of a PCSK9 nucleic acid that contains one ormore SNPs will contain the appropriate base substitution, insertion,repeat or deletion such that the compound is fully complementary to thealtered PCSK9 nucleic acid sequence.

A subgroup of these antisense compounds were selected for furthercharacterization, based on the ability of the selected antisensecompounds to inhibit PCSK9 expression selectively and effectively incell culture assays, set forth in the Examples. The antisense compoundsselected for further evaluation are listed in Table 1. The 5′ and 3′boundaries of the PCSK9 target sequence are shown for each antisensecompound, with reference to the nucleotide positions of SEQ ID NO: 1.

“Motif” in Table 1 refers to the use of chemically modified nucleosidesat the 5′ and 3′ ends of the antisense compounds. In a “5-10-5” motif,for example, five chemical modified nucleosides at both the 5′ and 3′ends flank ten unmodified nucleosides in the center of the antisensecompound. As disclosed in more detail below, modified nucleosides maymake the antisense compounds more resistant to nucleases, among otherthings, which generally improves the pharmacodynamic and pharmacokineticproperties of the antisense compounds when administered to anindividual.

Table 1 further discloses the nucleobase sequence of the antisensecompounds. In certain embodiments, an antisense compound has anucleobase sequence that, when written in the 5′ to 3′ direction,comprises the reverse complement of the target segment of a targetnucleic acid to which it is targeted.

TABLE 1 5′ Target 3′ Target SEQ ID Isis No. Site Site Sequence 5′-3′Motif NO 405881 1005 1024 CACCCTTGGCCACGCCGGCA 5-10-5 250 399819 25092528 CCCACTCAAGGGCCAGGCCA 5-10-5 65 395165 1004 1023ACCCTTGGCCACGCCGGCAT 5-10-5 28 405879 1002 1021 CCTTGGCCACGCCGGCATCC5-10-5 248 406008 602 621 CTTGGTGAGGTATCCCCGGC 5-10-5 188 405891 15671586 TCCTCAGGGAACCAGGCCTC 5-10-5 352 395186 2100 2119CTTTGCATTCCAGACCTGGG 5-10-5 60 405988 1854 1873 GGCAGCACCTGGCAATGGCG5-10-5 381 405994 1928 1947 GCAGTGGACACGGGTCCCCA 5-10-5 400 406023 787806 TGGTATTCATCCGCCCGGTA 5-10-5 212 395187 2310 2329GGCAGCAGATGGCAACGGCT 5-10-5 62 395185 1920 1939 CACGGGTCCCCATGCTGGCC5-10-5 59 406033 967 986 CCTGCCAGGTGGGTGCCATG 5-10-5 237 405923 12951314 GGCATTGGTGGCCCCAACTG 5-10-5 288 399900 1569 1588GGTCCTCAGGGAACCAGGCC 3-14-3 50 405995 1930 1949 TGGCAGTGGACACGGGTCCC5-10-5 402 405991 1922 1941 GACACGGGTCCCCATGCTGG 5-10-5 394 406005 559578 CGGGCAGTGCGCTCTGACTG 5-10-5 180 399793 417 436 CCTCGGAACGCAAGGCTAGC5-10-5 8 395152 410 429 ACGCAAGGCTAGCACCAGCT 5-10-5 7

Antisense Compound Motifs

In certain embodiments, antisense compounds targeted to a PCSK9 nucleicacid have chemically modified subunits arranged in patterns, or motifs,to confer to the antisense compounds properties such as enhanced theinhibitory activity, increased binding affinity for a target nucleicacid, or resistance to degradation by in vivo nucleases.

Chimeric antisense compounds typically contain at least one regionmodified so as to confer increased resistance to nuclease degradation,increased cellular uptake, increased binding affinity for the targetnucleic acid, and/or increased inhibitory activity. A second region of achimeric antisense compound may serve as a substrate for the cellularendonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.

Antisense compounds having a gapmer motif are considered chimericantisense compounds. In a gapmer an internal position having a pluralityof nucleotides that supports RNaseH cleavage is positioned betweenexternal regions having a plurality of nucleotides that are chemicallydistinct from the nucleosides of the internal region. In the case of anantisense oligonucleotide having a gapmer motif, the gap segmentgenerally serves as the substrate for endonuclease cleavage, while thewing segments comprise modified nucleosides. The regions of a gapmer aredifferentiated by the types of sugar moieties comprising each distinctregion. The types of sugar moieties that are used to differentiate theregions of a gapmer may in some embodiments include β-D-ribonucleosides,β-D-deoxyribonucleosides, 2′-modified nucleosides (such 2′-modifiednucleosides may include 2′-MOE, and 2′-O—CH₃, among others), andbicyclic sugar modified nucleosides (such bicyclic sugar modifiednucleosides may include those having a 4′-(CH2)n-O-2′ bridge, where n=1or n=2). In general, each distinct region comprises uniform sugarmoieties. The wing-gap-wing motif is frequently described as “X—Y—Z”,where “X” represents the length of the 5′ wing region, “Y” representsthe length of the gap region, and “Z” represents the length of the 3′wing region.

In some embodiments, an antisense compound targeted to a PCSK9 nucleicacid has a gap-widened motif. In other embodiments, an antisenseoligonucleotide targeted to a PCSK9 nucleic acid has a gap-widenedmotif.

The gap-widened antisense oligonucleotides described herein may havevarious wing-gap-wing motifs selected from: 1-16-1, 2-15-1, 1-15-2,1-14-3, 3-14-1, 2-14-2, 1-13-4, 4-13-1, 2-13-3, 3-13-2, 1-12-5, 5-12-1,2-12-4, 4-12-2, 3-12-3, 1-11-6, 6-11-1, 2-11-5, 5-11-2, 3-11-4, 4-11-3,1-17-1, 2-16-1, 1-16-2, 1-15-3, 3-15-1, 2-15-2, 1-14-4, 4-14-1, 2-14-3,3-14-2, 1-13-5, 5-13-1, 2-13-4, 4-13-2, 3-13-3, 1-12-6, 6-12-1, 2-12-5,5-12-2, 3-12-4, 4-12-3, 1-11-7, 7-11-1, 2-11-6, 6-11-2, 3-11-5, 5-11-3,4-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 1-16-3, 2-16-2, 1-15-4, 4-15-1,2-15-3, 3-15-2, 1-14-5, 5-14-1, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1,2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5,5-12-3, 4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5,5-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 3-16-1, 2-16-2, 1-15-4, 4-15-1,2-15-3, 3-15-2, 1-14-5, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5,5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3,4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4,1-19-1, 1-18-2, 2-18-1, 1-17-3, 3-17-1, 2-17-2, 1-16-4, 4-16-1, 2-16-3,3-16-2, 1-15-5, 2-15-4, 4-15-2, 3-15-3, 1-14-6, 6-14-1, 2-14-5, 5-14-2,3-14-4, 4-14-3, 1-13-7, 7-13-1, 2-13-6, 6-13-2, 3-13-5, 5-13-3, 4-13-4,1-12-8, 8-12-1, 2-12-7, 7-12-2, 3-12-6, 6-12-3, 4-12-5, 5-12-4, 2-11-8,8-11-2, 3-11-7, 7-11-3, 4-11-6, 6-11-4, 5-11-5, 1-20-1, 1-19-2, 2-19-1,1-18-3, 3-18-1, 2-18-2, 1-17-4, 4-17-1, 2-17-3, 3-17-2, 1-16-5, 2-16-4,4-16-2, 3-16-3, 1-15-6, 6-15-1, 2-15-5, 5-15-2, 3-15-4, 4-15-3, 1-14-7,7-14-1, 2-14-6, 6-14-2, 3-14-5, 5-14-3, 4-14-4, 1-13-8, 8-13-1, 2-13-7,7-13-2, 3-13-6, 6-13-3, 4-13-5, 5-13-4, 2-12-8, 8-12-2, 3-12-7, 7-12-3,4-12-6, 6-12-4, 5-12-5, 3-11-8, 8-11-3, 4-11-7, 7-11-4, 5-11-6, 6-11-5,1-21-1, 1-20-2, 2-20-1, 1-20-3, 3-19-1, 2-19-2, 1-18-4, 4-18-1, 2-18-3,3-18-2, 1-17-5, 2-17-4, 4-17-2, 3-17-3, 1-16-6, 6-16-1, 2-16-5, 5-16-2,3-16-4, 4-16-3, 1-15-7, 7-15-1, 2-15-6, 6-15-2, 3-15-5, 5-15-3, 4-15-4,1-14-8, 8-14-1, 2-14-7, 7-14-2, 3-14-6, 6-14-3, 4-14-5, 5-14-4, 2-13-8,8-13-2, 3-13-7, 7-13-3, 4-13-6, 6-13-4, 5-13-5, 1-12-10, 10-12-1,2-12-9, 9-12-2, 3-12-8, 8-12-3, 4-12-7, 7-12-4, 5-12-6, 6-12-5, 4-11-8,8-11-4, 5-11-7, 7-11-5, 6-11-6, 1-22-1, 1-21-2, 2-21-1, 1-21-3, 3-20-1,2-20-2, 1-19-4, 4-19-1, 2-19-3, 3-19-2, 1-18-5, 2-18-4, 4-18-2, 3-18-3,1-17-6, 6-17-1, 2-17-5, 5-17-2, 3-17-4, 4-17-3, 1-16-7, 7-16-1, 2-16-6,6-16-2, 3-16-5, 5-16-3, 4-16-4, 1-15-8, 8-15-1, 2-15-7, 7-15-2, 3-15-6,6-15-3, 4-15-5, 5-15-4, 2-14-8, 8-14-2, 3-14-7, 7-14-3, 4-14-6, 6-14-4,5-14-5, 3-13-8, 8-13-3, 4-13-7, 7-13-4, 5-13-6, 6-13-5, 4-12-8, 8-12-4,5-12-7, 7-12-5, 6-12-6, 5-11-8, 8-11-5, 6-11-7, or 7-11-6. In certainpreferred embodiments, a gap-widened motif includes, but is not limitedto, 2-13-5, 3-14-3, 3-14-4 gapmer motif.

In one embodiment, a gap-widened antisense oligonucleotide targeted to aPCSK9 nucleic acid has a gap segment of fourteen 2′-deoxyribonucleotidespositioned between wing segments of three chemically modifiednucleosides. In one embodiment, the chemical modification comprises a2′-sugar modification. In another embodiment, the chemical modificationcomprises a 2′-MOE sugar modification.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acidpossess a 5-10-5 gapmer motif.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

Nucleotide sequences that encode PCSK9 include, without limitation, thefollowing: GENBANK® Accession No. NM_174936.2, first deposited withGENBANK® on Jun. 1, 2003, and incorporated herein as SEQ ID NO: 1;nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_032977.8,first deposited with GENBANK® on Feb. 26, 2006, and incorporated hereinas SEQ ID NO: 2; and GENBANK® Accession No. AK124635.1, first depositedwith GENBANK® on Sep. 8, 2003, and incorporated herein as SEQ ID NO: 3.

It is noted that some portions of these nucleotide sequences shareidentical sequence. For example, portions of SEQ ID NO: 1 are identicalto portions of SEQ ID NO: 2; portions of SEQ ID NO: 1 are identical toportions of SEQ ID NO: 3; and portions of SEQ ID NO: 2 are identical toportions of SEQ ID NO: 3. Accordingly, antisense compounds targeted toSEQ ID NO: 1 may also target SEQ ID NO: 2 and/or SEQ ID NO: 3; antisensecompounds targeted to SEQ ID NO: 2 may also target SEQ ID NO: 1 and/orSEQ ID NO: 3; and antisense compounds targeted to SEQ ID NO: 3 may alsotarget SEQ ID NO: 1 and/or SEQ ID NO: 2. Examples of such antisensecompounds are shown in the following tables.

In certain embodiments, antisense compounds target a PCSK9 nucleic acidhaving the sequence of GENBANK® Accession No. NM_174936.2, firstdeposited with GENBANK® on Jun. 1, 2003, and incorporated herein as SEQID NO: 1. In certain such embodiments, an antisense oligonucleotidetargets SEQ ID NO: 1. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 1 is at least 90%complementary to SEQ ID NO: 1. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 1 is at least 95%complementary to SEQ ID NO: 1. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 1 is 100% complementaryto SEQ ID NO: 1. In certain embodiments, an antisense oligonucleotidetargeted to SEQ ID NO: 1 comprises a nucleotide sequence selected fromthe nucleotide sequences set forth in Table 2.

TABLE 2 Nucleotide sequences targeted to NM_174936.2 (SEQ ID NO: 1)5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID NO NO: 1 NO: 1Sequence (5′-3′) 4 135 154 GCGCGGAATCCTGGCTGGGA 5 242 261GAGGAGACCTAGAGGCCGTG 159 294 313 GCCTGGAGCTGACGGTGCCC 160 298 317GACCGCCTGGAGCTGACGGT 6 300 319 AGGACCGCCTGGAGCTGACG 162 406 425AAGGCTAGCACCAGCTCCTC 163 407 426 CAAGGCTAGCACCAGCTCCT 164 408 427GCAAGGCTAGCACCAGCTCC 165 409 428 CGCAAGGCTAGCACCAGCTC 7 410 429ACGCAAGGCTAGCACCAGCT 166 411 430 AACGCAAGGCTAGCACCAGC 167 412 431GAACGCAAGGCTAGCACCAG 168 413 432 GGAACGCAAGGCTAGCACCA 169 414 433CGGAACGCAAGGCTAGCACC 8 417 436 CCTCGGAACGCAAGGCTAGC 170 421 440TCCTCCTCGGAACGCAAGGC 171 446 465 GTGCTCGGGTGCTTCGGCCA 172 466 485TGGAAGGTGGCTGTGGTTCC 9 480 499 CCTTGGCGCAGCGGTGGAAG 173 482 501ATCCTTGGCGCAGCGGTGGA 174 484 503 GGATCCTTGGCGCAGCGGTG 175 488 507CCACGGATCCTTGGCGCAGC 176 507 526 CGTAGGTGCCAGGCAACCTC 177 545 564TGACTGCGAGAGGTGGGTCT 178 555 574 CAGTGCGCTCTGACTGCGAG 179 557 576GGCAGTGCGCTCTGACTGCG 180 559 578 CGGGCAGTGCGCTCTGACTG 10 561 580GGCGGGCAGTGCGCTCTGAC 181 562 581 CGGCGGGCAGTGCGCTCTGA 182 591 610ATCCCCGGCGGGCAGCCTGG 183 595 614 AGGTATCCCCGGCGGGCAGC 184 597 616TGAGGTATCCCCGGCGGGCA 185 598 617 GTGAGGTATCCCCGGCGGGC 186 599 618GGTGAGGTATCCCCGGCGGG 11 600 619 TGGTGAGGTATCCCCGGCGG 187 601 620TTGGTGAGGTATCCCCGGCG 188 602 621 CTTGGTGAGGTATCCCCGGC 189 603 622TCTTGGTGAGGTATCCCCGG 190 604 623 ATCTTGGTGAGGTATCCCCG 191 605 624GATCTTGGTGAGGTATCCCC 12 606 625 GGATCTTGGTGAGGTATCCC 192 607 626AGGATCTTGGTGAGGTATCC 193 609 628 GCAGGATCTTGGTGAGGTAT 194 611 630ATGCAGGATCTTGGTGAGGT 195 613 632 ACATGCAGGATCTTGGTGAG 13 615 634AGACATGCAGGATCTTGGTG 196 617 636 GAAGACATGCAGGATCTTGG 14 620 639ATGGAAGACATGCAGGATCT 197 628 647 AGAAGGCCATGGAAGACATG 198 638 657GAAGCCAGGAAGAAGGCCAT 15 646 665 TTCACCAGGAAGCCAGGAAG 199 648 667TCTTCACCAGGAAGCCAGGA 16 651 670 TCATCTTCACCAGGAAGCCA 200 653 672ACTCATCTTCACCAGGAAGC 201 655 674 CCACTCATCTTCACCAGGAA 202 657 676CGCCACTCATCTTCACCAGG 203 659 678 GTCGCCACTCATCTTCACCA 204 661 680AGGTCGCCACTCATCTTCAC 205 663 682 GCAGGTCGCCACTCATCTTC 206 665 684CAGCAGGTCGCCACTCATCT 207 667 686 TCCAGCAGGTCGCCACTCAT 208 685 704GGCAACTTCAAGGCCAGCTC 17 705 724 CCTCGATGTAGTCGACATGG 209 724 743GCAAAGACAGAGGAGTCCTC 210 782 801 TTCATCCGCCCGGTACCGTG 211 784 803TATTCATCCGCCCGGTACCG 18 785 804 GTATTCATCCGCCCGGTACC 212 787 806TGGTATTCATCCGCCCGGTA 213 789 808 GCTGGTATTCATCCGCCCGG 214 791 810GGGCTGGTATTCATCCGCCC 215 821 840 ATACACCTCCACCAGGCTGC 216 832 851GTGTCTAGGAGATACACCTC 19 835 854 CTGGTGTCTAGGAGATACAC 217 837 856TGCTGGTGTCTAGGAGATAC 20 840 859 GTATGCTGGTGTCTAGGAGA 21 860 879GATTTCCCGGTGGTCACTCT 218 862 881 TCGATTTCCCGGTGGTCACT 219 863 882CTCGATTTCCCGGTGGTCAC 220 864 883 CCTCGATTTCCCGGTGGTCA 221 865 884CCCTCGATTTCCCGGTGGTC 22 866 885 GCCCTCGATTTCCCGGTGGT 222 867 886TGCCCTCGATTTCCCGGTGG 223 868 887 CTGCCCTCGATTTCCCGGTG 224 869 888CCTGCCCTCGATTTCCCGGT 225 870 889 CCCTGCCCTCGATTTCCCGG 226 874 893ATGACCCTGCCCTCGATTTC 227 876 895 CCATGACCCTGCCCTCGATT 228 878 897GACCATGACCCTGCCCTCGA 23 880 899 GTGACCATGACCCTGCCCTC 229 882 901CGGTGACCATGACCCTGCCC 230 884 903 GTCGGTGACCATGACCCTGC 231 886 905AAGTCGGTGACCATGACCCT 232 888 907 CGAAGTCGGTGACCATGACC 24 890 909CTCGAAGTCGGTGACCATGA 233 898 917 GGCACATTCTCGAAGTCGGT 25 923 942GTGGAAGCGGGTCCCGTCCT 234 933 952 TGGCCTGTCTGTGGAAGCGG 235 960 979GGTGGGTGCCATGACTGTCA 236 963 982 CCAGGTGGGTGCCATGACTG 237 967 986CCTGCCAGGTGGGTGCCATG 26 970 989 ACCCCTGCCAGGTGGGTGCC 238 972 991CCACCCCTGCCAGGTGGGTG 27 975 994 TGACCACCCCTGCCAGGTGG 239 977 996GCTGACCACCCCTGCCAGGT 240 985 1004 TCCCGGCCGCTGACCACCCC 241 989 1008GGCATCCCGGCCGCTGACCA 242 992 1011 GCCGGCATCCCGGCCGCTGA 243 997 1016GCCACGCCGGCATCCCGGCC 244 998 1017 GGCCACGCCGGCATCCCGGC 245 999 1018TGGCCACGCCGGCATCCCGG 246 1000 1019 TTGGCCACGCCGGCATCCCG 247 1001 1020CTTGGCCACGCCGGCATCCC 248 1002 1021 CCTTGGCCACGCCGGCATCC 249 1003 1022CCCTTGGCCACGCCGGCATC 28 1004 1023 ACCCTTGGCCACGCCGGCAT 447 1004 1023ACCCTTGGTCACGCCGGCAT 250 1005 1024 CACCCTTGGCCACGCCGGCA 251 1006 1025GCACCCTTGGCCACGCCGGC 252 1007 1026 GGCACCCTTGGCCACGCCGG 253 1008 1027TGGCACCCTTGGCCACGCCG 254 1009 1028 CTGGCACCCTTGGCCACGCC 255 1010 1029GCTGGCACCCTTGGCCACGC 256 1015 1034 CGCATGCTGGCACCCTTGGC 257 1036 1055CAGTTGAGCACGCGCAGGCT 258 1038 1057 GGCAGTTGAGCACGCGCAGG 29 1040 1059TTGGCAGTTGAGCACGCGCA 259 1042 1061 CCTTGGCAGTTGAGCACGCG 30 1045 1064TTCCCTTGGCAGTTGAGCAC 260 1047 1066 CCTTCCCTTGGCAGTTGAGC 261 1051 1070GTGCCCTTCCCTTGGCAGTT 262 1053 1072 CCGTGCCCTTCCCTTGGCAG 263 1064 1083GGTGCCGCTAACCGTGCCCT 264 1076 1095 CAGGCCTATGAGGGTGCCGC 31 1077 1096CCAGGCCTATGAGGGTGCCG 458 1079 1092 GCCTATGAGGGTGC 459 1084 1097TCCAGGCCTATGAG 265 1088 1107 CCGAATAAACTCCAGGCCTA 266 1096 1115TGGCTTTTCCGAATAAACTC 32 1098 1117 GCTGGCTTTTCCGAATAAAC 267 1100 1119CAGCTGGCTTTTCCGAATAA 268 1102 1121 ACCAGCTGGCTTTTCCGAAT 269 1104 1123GGACCAGCTGGCTTTTCCGA 270 1108 1127 GGCTGGACCAGCTGGCTTTT 271 1119 1138GTGGCCCCACAGGCTGGACC 272 1132 1151 AGCAGCACCACCAGTGGCCC 273 1154 1173GCTGTACCCACCCGCCAGGG 274 1200 1219 CGACCCCAGCCCTCGCCAGG 33 1210 1229GTGACCAGCACGACCCCAGC 275 1212 1231 CGGTGACCAGCACGACCCCA 276 1214 1233AGCGGTGACCAGCACGACCC 277 1216 1235 GCAGCGGTGACCAGCACGAC 278 1218 1237CGGCAGCGGTGACCAGCACG 279 1219 1238 CCGGCAGCGGTGACCAGCAC 280 1222 1241TTGCCGGCAGCGGTGACCAG 281 1224 1243 AGTTGCCGGCAGCGGTGACC 282 1226 1245GAAGTTGCCGGCAGCGGTGA 283 1228 1247 CGGAAGTTGCCGGCAGCGGT 284 1230 1249CCCGGAAGTTGCCGGCAGCG 285 1232 1251 GTCCCGGAAGTTGCCGGCAG 286 1273 1292ATGACCTCGGGAGCTGAGGC 287 1283 1302 CCCAACTGTGATGACCTCGG 288 1295 1314GGCATTGGTGGCCCCAACTG 149 1297 1316 TGGGCATTGGTGGCCCCAAC 289 1305 1324GCTGGTCTTGGGCATTGGTG 290 1318 1337 CCCAGGGTCACCGGCTGGTC 291 1320 1339TCCCCAGGGTCACCGGCTGG 292 1322 1341 AGTCCCCAGGGTCACCGGCT 293 1324 1343AAAGTCCCCAGGGTCACCGG 34 1326 1345 CCAAAGTCCCCAGGGTCACC 294 1328 1347CCCCAAAGTCCCCAGGGTCA 128 1330 1349 GTCCCCAAAGTCCCCAGGGT 295 1333 1352TTGGTCCCCAAAGTCCCCAG 35 1335 1354 AGTTGGTCCCCAAAGTCCCC 296 1337 1356AAAGTTGGTCCCCAAAGTCC 36 1340 1359 GCCAAAGTTGGTCCCCAAAG 297 1342 1361CGGCCAAAGTTGGTCCCCAA 298 1344 1363 AGCGGCCAAAGTTGGTCCCC 299 1346 1365ACAGCGGCCAAAGTTGGTCC 300 1348 1367 ACACAGCGGCCAAAGTTGGT 301 1350 1369CCACACAGCGGCCAAAGTTG 37 1352 1371 GTCCACACAGCGGCCAAAGT 302 1354 1373AGGTCCACACAGCGGCCAAA 303 1356 1375 AGAGGTCCACACAGCGGCCA 304 1358 1377AAAGAGGTCCACACAGCGGC 38 1361 1380 GGCAAAGAGGTCCACACAGC 305 1380 1399CAATGATGTCCTCCCCTGGG 306 1387 1406 GAGGCACCAATGATGTCCTC 39 1389 1408TGGAGGCACCAATGATGTCC 307 1391 1410 GCTGGAGGCACCAATGATGT 308 1393 1412TCGCTGGAGGCACCAATGAT 309 1395 1414 AGTCGCTGGAGGCACCAATG 310 1397 1416GCAGTCGCTGGAGGCACCAA 40 1400 1419 GCTGCAGTCGCTGGAGGCAC 311 1402 1421GTGCTGCAGTCGCTGGAGGC 312 1404 1423 AGGTGCTGCAGTCGCTGGAG 313 1406 1425GCAGGTGCTGCAGTCGCTGG 314 1407 1426 AGCAGGTGCTGCAGTCGCTG 315 1409 1428AAAGCAGGTGCTGCAGTCGC 41 1411 1430 ACAAAGCAGGTGCTGCAGTC 316 1413 1432ACACAAAGCAGGTGCTGCAG 317 1415 1434 TGACACAAAGCAGGTGCTGC 318 1425 1444TCCCACTCTGTGACACAAAG 101 1465 1484 ATGGCTGCAATGCCAGCCAC 319 1467 1486TCATGGCTGCAATGCCAGCC 42 1470 1489 GCATCATGGCTGCAATGCCA 320 1472 1491CAGCATCATGGCTGCAATGC 321 1474 1493 GACAGCATCATGGCTGCAAT 322 1476 1495CAGACAGCATCATGGCTGCA 43 1478 1497 GGCAGACAGCATCATGGCTG 323 1480 1499TCGGCAGACAGCATCATGGC 324 1482 1501 GCTCGGCAGACAGCATCATG 325 1484 1503CGGCTCGGCAGACAGCATCA 326 1486 1505 TCCGGCTCGGCAGACAGCAT 327 1500 1519CGGCCAGGGTGAGCTCCGGC 328 1513 1532 CTCTGCCTCAACTCGGCCAG 329 1515 1534GTCTCTGCCTCAACTCGGCC 330 1517 1536 CAGTCTCTGCCTCAACTCGG 331 1519 1538ATCAGTCTCTGCCTCAACTC 332 1521 1540 GGATCAGTCTCTGCCTCAAC 333 1523 1542GTGGATCAGTCTCTGCCTCA 334 1525 1544 AAGTGGATCAGTCTCTGCCT 44 1526 1545GAAGTGGATCAGTCTCTGCC 335 1528 1547 GAGAAGTGGATCAGTCTCTG 336 1530 1549CAGAGAAGTGGATCAGTCTC 337 1532 1551 GGCAGAGAAGTGGATCAGTC 45 1534 1553TTGGCAGAGAAGTGGATCAG 338 1536 1555 CTTTGGCAGAGAAGTGGATC 46 1539 1558CATCTTTGGCAGAGAAGTGG 339 1541 1560 GACATCTTTGGCAGAGAAGT 340 1543 1562ATGACATCTTTGGCAGAGAA 47 1545 1564 TGATGACATCTTTGGCAGAG 341 1547 1566ATTGATGACATCTTTGGCAG 342 1549 1568 TCATTGATGACATCTTTGGC 48 1552 1571GCCTCATTGATGACATCTTT 343 1554 1573 AGGCCTCATTGATGACATCT 344 1556 1575CCAGGCCTCATTGATGACAT 345 1558 1577 AACCAGGCCTCATTGATGAC 346 1560 1579GGAACCAGGCCTCATTGATG 347 1561 1580 GGGAACCAGGCCTCATTGAT 348 1562 1581AGGGAACCAGGCCTCATTGA 349 1563 1582 CAGGGAACCAGGCCTCATTG 49 1564 1583TCAGGGAACCAGGCCTCATT 49 1564 1583 TCAGGGAACCAGGCCTCATT 350 1565 1584CTCAGGGAACCAGGCCTCAT 351 1566 1585 CCTCAGGGAACCAGGCCTCA 352 1567 1586TCCTCAGGGAACCAGGCCTC 353 1568 1587 GTCCTCAGGGAACCAGGCCT 50 1569 1588GGTCCTCAGGGAACCAGGCC 354 1570 1589 TGGTCCTCAGGGAACCAGGC 355 1571 1590CTGGTCCTCAGGGAACCAGG 356 1572 1591 GCTGGTCCTCAGGGAACCAG 357 1573 1592CGCTGGTCCTCAGGGAACCA 87 1576 1595 ACCCGCTGGTCCTCAGGGAA 358 1578 1597GTACCCGCTGGTCCTCAGGG 359 1580 1599 CAGTACCCGCTGGTCCTCAG 51 1583 1602GGTCAGTACCCGCTGGTCCT 119 1605 1624 GCAGGGCGGCCACCAGGTTG 360 1628 1647ACCTGCCCCATGGGTGCTGG 52 1640 1659 AAACAGCTGCCAACCTGCCC 361 1642 1661CAAAACAGCTGCCAACCTGC 53 1645 1664 CTGCAAAACAGCTGCCAACC 362 1647 1666TCCTGCAAAACAGCTGCCAA 363 1649 1668 AGTCCTGCAAAACAGCTGCC 364 1660 1679GCTGACCATACAGTCCTGCA 365 1672 1691 GGCCCCGAGTGTGCTGACCA 54 1675 1694GTAGGCCCCGAGTGTGCTGA 366 1677 1696 GTGTAGGCCCCGAGTGTGCT 367 1679 1698CCGTGTAGGCCCCGAGTGTG 368 1681 1700 ATCCGTGTAGGCCCCGAGTG 369 1683 1702CCATCCGTGTAGGCCCCGAG 370 1685 1704 GGCCATCCGTGTAGGCCCCG 371 1687 1706GTGGCCATCCGTGTAGGCCC 372 1735 1754 CTGGAGCAGCTCAGCAGCTC 460 1735 1748CAGCTCAGCAGCTC 373 1737 1756 AACTGGAGCAGCTCAGCAGC 55 1740 1759AGAAACTGGAGCAGCTCAGC 374 1742 1761 GGAGAAACTGGAGCAGCTCA 375 1744 1763CTGGAGAAACTGGAGCAGCT 56 1746 1765 TCCTGGAGAAACTGGAGCAG 57 1812 1831CGTTGTGGGCCCGGCAGACC 376 1849 1868 CACCTGGCAATGGCGTAGAC 377 1850 1869GCACCTGGCAATGGCGTAGA 378 1851 1870 AGCACCTGGCAATGGCGTAG 379 1852 1871CAGCACCTGGCAATGGCGTA 380 1853 1872 GCAGCACCTGGCAATGGCGT 381 1854 1873GGCAGCACCTGGCAATGGCG 382 1855 1874 AGGCAGCACCTGGCAATGGC 383 1856 1875CAGGCAGCACCTGGCAATGG 384 1857 1876 GCAGGCAGCACCTGGCAATG 58 1858 1877AGCAGGCAGCACCTGGCAAT 385 1859 1878 TAGCAGGCAGCACCTGGCAA 386 1860 1879GTAGCAGGCAGCACCTGGCA 387 1905 1924 TGGCCTCAGCTGGTGGAGCT 388 1915 1934GTCCCCATGCTGGCCTCAGC 389 1916 1935 GGTCCCCATGCTGGCCTCAG 390 1917 1936GGGTCCCCATGCTGGCCTCA 391 1918 1937 CGGGTCCCCATGCTGGCCTC 392 1919 1938ACGGGTCCCCATGCTGGCCT 59 1920 1939 CACGGGTCCCCATGCTGGCC 59 1920 1939CACGGGTCCCCATGCTGGCC 393 1921 1940 ACACGGGTCCCCATGCTGGC 394 1922 1941GACACGGGTCCCCATGCTGG 395 1923 1942 GGACACGGGTCCCCATGCTG 396 1924 1943TGGACACGGGTCCCCATGCT 397 1925 1944 GTGGACACGGGTCCCCATGC 398 1926 1945AGTGGACACGGGTCCCCATG 399 1927 1946 CAGTGGACACGGGTCCCCAT 400 1928 1947GCAGTGGACACGGGTCCCCA 401 1929 1948 GGCAGTGGACACGGGTCCCC 402 1930 1949TGGCAGTGGACACGGGTCCC 403 1931 1950 GTGGCAGTGGACACGGGTCC 404 1932 1951GGTGGCAGTGGACACGGGTC 405 1933 1952 TGGTGGCAGTGGACACGGGT 406 1936 1955TGTTGGTGGCAGTGGACACG 407 1962 1981 AGCTGCAGCCTGTGAGGACG 408 1990 2009GTGCCAAGGTCCTCCACCTC 409 2010 2029 TCAGCACAGGCGGCTTGTGG 410 2040 2059CCACGCACTGGTTGGGCTGA 60 2100 2119 CTTTGCATTCCAGACCTGGG 411 2101 2120ACTTTGCATTCCAGACCTGG 412 2102 2121 GACTTTGCATTCCAGACCTG 413 2103 2122TGACTTTGCATTCCAGACCT 414 2104 2123 TTGACTTTGCATTCCAGACC 61 2105 2124CTTGACTTTGCATTCCAGAC 415 2107 2126 TCCTTGACTTTGCATTCCAG 416 2120 2139CGGGATTCCATGCTCCTTGA 417 2150 2169 GCAGGCCACGGTCACCTGCT 418 2187 2206GGAGGGCACTGCAGCCAGTC 419 2305 2324 CAGATGGCAACGGCTGTCAC 420 2306 2325GCAGATGGCAACGGCTGTCA 421 2307 2326 AGCAGATGGCAACGGCTGTC 422 2308 2327CAGCAGATGGCAACGGCTGT 423 2309 2328 GCAGCAGATGGCAACGGCTG 62 2310 2329GGCAGCAGATGGCAACGGCT 424 2311 2330 CGGCAGCAGATGGCAACGGC 425 2312 2331CCGGCAGCAGATGGCAACGG 426 2313 2332 TCCGGCAGCAGATGGCAACG 427 2314 2333CTCCGGCAGCAGATGGCAAC 428 2315 2334 GCTCCGGCAGCAGATGGCAA 429 2325 2344CCAGGTGCCGGCTCCGGCAG 430 2335 2354 GAGGCCTGCGCCAGGTGCCG 154 2410 2429TTTTAAAGCTCAGCCCCAGC 63 2415 2434 AACCATTTTAAAGCTCAGCC 64 2504 2523TCAAGGGCCAGGCCAGCAGC 65 2509 2528 CCCACTCAAGGGCCAGGCCA 122 2582 2601GGAGGGAGCTTCCTGGCACC 66 2597 2616 ATGCCCCACAGTGAGGGAGG 67 2606 2625AATGGTGAAATGCCCCACAG 153 2649 2668 TTGGGAGCAGCTGGCAGCAC 68 2750 2769CATGGGAAGAATCCTGCCTC 431 2828 2847 ATGAGGGCCATCAGCACCTT 432 2829 2848GATGAGGGCCATCAGCACCT 433 2830 2849 AGATGAGGGCCATCAGCACC 434 2831 2850GAGATGAGGGCCATCAGCAC 69 2832 2851 GGAGATGAGGGCCATCAGCA 435 2833 2852TGGAGATGAGGGCCATCAGC 436 2834 2853 CTGGAGATGAGGGCCATCAG 437 2835 2854GCTGGAGATGAGGGCCATCA 438 2836 2855 AGCTGGAGATGAGGGCCATC 461 2877 2890TTAATCAGGGAGCC 70 2900 2919 TAGATGCCATCCAGAAAGCT 439 2902 2921GCTAGATGCCATCCAGAAAG 440 2903 2922 GGCTAGATGCCATCCAGAAA 441 2904 2923TGGCTAGATGCCATCCAGAA 442 2905 2924 CTGGCTAGATGCCATCCAGA 71 2906 2925TCTGGCTAGATGCCATCCAG 443 2907 2926 CTCTGGCTAGATGCCATCCA 444 2908 2927CCTCTGGCTAGATGCCATCC 445 2909 2928 GCCTCTGGCTAGATGCCATC 446 2910 2929AGCCTCTGGCTAGATGCCAT 72 2983 3002 GGCATAGAGCAGAGTAAAGG 73 2988 3007AGCCTGGCATAGAGCAGAGT 135 2994 3013 TAGCACAGCCTGGCATAGAG 112 3227 3246GAAGAGGCTTGGCTTCAGAG 74 3233 3252 AAGTAAGAAGAGGCTTGGCT 75 3437 3456GCTCAAGGAGGGACAGTTGT 76 3472 3491 AAAGATAAATGTCTGCTTGC 77 3477 3496ACCCAAAAGATAAATGTCTG 78 3543 3562 TCTTCAAGTTACAAAAGCAA 99 3550 3569ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to aPCSK9 nucleic acid. In certain such embodiments, gapmer antisensecompounds are targeted to SEQ ID NO: 1. In certain such embodiments, thenucleotide sequences illustrated in Table 2 have a 5-10-5 gapmer motif.Table 3 illustrates gapmer antisense compounds targeted to SEQ ID NO: 1,having a 5-10-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 3 Gapmer antisense compounds having a 5-10-5 motif targeted to SEQID NO: 1 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID IsisNo Motif NO NO: 1 NO: 1 Mismatches Sequence (5′-3′) 395149 5-10-5 4 135154 0 GCGCGGAATCCTGGCTGGGA 395150 5-10-5 5 242 261 0GAGGAGACCTAGAGGCCGTG 395151 5-10-5 6 300 319 0 AGGACCGCCTGGAGCTGACG395152 5-10-5 7 410 429 0 ACGCAAGGCTAGCACCAGCT 399793 5-10-5 8 417 436 0CCTCGGAACGCAAGGCTAGC 395153 5-10-5 9 480 499 0 CCTTGGCGCAGCGGTGGAAG395154 5-10-5 10 561 580 0 GGCGGGCAGTGCGCTCTGAC 395155 5-10-5 11 600 6190 TGGTGAGGTATCCCCGGCGG 399794 5-10-5 12 606 625 0 GGATCTTGGTGAGGTATCCC399795 5-10-5 13 615 634 0 AGACATGCAGGATCTTGGTG 395156 5-10-5 14 620 6390 ATGGAAGACATGCAGGATCT 395157 5-10-5 15 646 665 0 TTCACCAGGAAGCCAGGAAG399796 5-10-5 16 651 670 0 TCATCTTCACCAGGAAGCCA 395158 5-10-5 17 705 7240 CCTCGATGTAGTCGACATGG 395159 5-10-5 18 785 804 0 GTATTCATCCGCCCGGTACC395160 5-10-5 19 835 854 0 CTGGTGTCTAGGAGATACAC 399797 5-10-5 20 840 8590 GTATGCTGGTGTCTAGGAGA 395161 5-10-5 21 860 879 0 GATTTCCCGGTGGTCACTCT399798 5-10-5 22 866 885 0 GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 880 8990 GTGACCATGACCCTGCCCTC 395162 5-10-5 24 890 909 0 CTCGAAGTCGGTGACCATGA395163 5-10-5 25 923 942 0 GTGGAAGCGGGTCCCGTCCT 395164 5-10-5 26 970 9890 ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 975 994 0 TGACCACCCCTGCCAGGTGG395165 5-10-5 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT 395166 5-10-5 29 10401059 0 TTGGCAGTTGAGCACGCGCA 399801 5-10-5 30 1045 1064 0TTCCCTTGGCAGTTGAGCAC 395167 5-10-5 31 1077 1096 0 CCAGGCCTATGAGGGTGCCG395168 5-10-5 32 1098 1117 0 GCTGGCTTTTCCGAATAAAC 395169 5-10-5 33 12101229 0 GTGACCAGCACGACCCCAGC 395170 5-10-5 149 1297 1316 0TGGGCATTGGTGGCCCCAAC 395171 5-10-5 34 1326 1345 0 CCAAAGTCCCCAGGGTCACC395172 5-10-5 128 1330 1349 0 GTCCCCAAAGTCCCCAGGGT 399802 5-10-5 35 13351354 0 AGTTGGTCCCCAAAGTCCCC 395173 5-10-5 36 1340 1359 0GCCAAAGTTGGTCCCCAAAG 399803 5-10-5 37 1352 1371 0 GTCCACACAGCGGCCAAAGT395174 5-10-5 38 1361 1380 0 GGCAAAGAGGTCCACACAGC 395175 5-10-5 39 13891408 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 1400 1419 0GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 1411 1430 0 ACAAAGCAGGTGCTGCAGTC395176 5-10-5 101 1465 1484 0 ATGGCTGCAATGCCAGCCAC 399806 5-10-5 42 14701489 0 GCATCATGGCTGCAATGCCA 399807 5-10-5 43 1478 1497 0GGCAGACAGCATCATGGCTG 399808 5-10-5 44 1526 1545 0 GAAGTGGATCAGTCTCTGCC395177 5-10-5 45 1534 1553 0 TTGGCAGAGAAGTGGATCAG 399809 5-10-5 46 15391558 0 CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 1545 1564 0TGATGACATCTTTGGCAGAG 399811 5-10-5 48 1552 1571 0 GCCTCATTGATGACATCTTT399812 5-10-5 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 395178 5-10-5 50 15691588 0 GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 1576 1595 0ACCCGCTGGTCCTCAGGGAA 399813 5-10-5 51 1583 1602 0 GGTCAGTACCCGCTGGTCCT395180 5-10-5 119 1605 1624 0 GCAGGGCGGCCACCAGGTTG 395181 5-10-5 52 16401659 0 AAACAGCTGCCAACCTGCCC 399814 5-10-5 53 1645 1664 0CTGCAAAACAGCTGCCAACC 395182 5-10-5 54 1675 1694 0 GTAGGCCCCGAGTGTGCTGA399815 5-10-5 55 1740 1759 0 AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 17461765 0 TCCTGGAGAAACTGGAGCAG 395183 5-10-5 57 1812 1831 0CGTTGTGGGCCCGGCAGACC 395184 5-10-5 58 1858 1877 0 AGCAGGCAGCACCTGGCAAT395185 5-10-5 59 1920 1939 0 CACGGGTCCCCATGCTGGCC 395186 5-10-5 60 21002119 0 CTTTGCATTCCAGACCTGGG 399817 5-10-5 61 2105 2124 0CTTGACTTTGCATTCCAGAC 395187 5-10-5 62 2310 2329 0 GGCAGCAGATGGCAACGGCT395188 5-10-5 154 2410 2429 0 TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 63 24152434 0 AACCATTTTAAAGCTCAGCC 395189 5-10-5 64 2504 2523 0TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 2509 2528 0 CCCACTCAAGGGCCAGGCCA399820 5-10-5 122 2582 2601 0 GGAGGGAGCTTCCTGGCACC 395190 5-10-5 66 25972616 0 ATGCCCCACAGTGAGGGAGG 395191 5-10-5 67 2606 2625 0AATGGTGAAATGCCCCACAG 395192 5-10-5 153 2649 2668 0 TTGGGAGCAGCTGGCAGCAC395193 5-10-5 68 2750 2769 0 CATGGGAAGAATCCTGCCTC 395194 5-10-5 69 28322851 0 GGAGATGAGGGCCATCAGCA 395195 5-10-5 70 2900 2919 0TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 2906 2925 0 TCTGGCTAGATGCCATCCAG395196 5-10-5 72 2983 3002 0 GGCATAGAGCAGAGTAAAGG 399822 5-10-5 73 29883007 0 AGCCTGGCATAGAGCAGAGT 399823 5-10-5 135 2994 3013 0TAGCACAGCCTGGCATAGAG 395197 5-10-5 112 3227 3246 0 GAAGAGGCTTGGCTTCAGAG399824 5-10-5 74 3233 3252 0 AAGTAAGAAGAGGCTTGGCT 395198 5-10-5 75 34373456 0 GCTCAAGGAGGGACAGTTGT 395199 5-10-5 76 3472 3491 0AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 3477 3496 0 ACCCAAAAGATAAATGTCTG395200 5-10-5 78 3543 3562 0 TCTTCAAGTTACAAAAGCAA 399826 5-10-5 99 35503569 0 ATAAATATCTTCAAGTTACA 405861 5-10-5 162 406 425 0AAGGCTAGCACCAGCTCCTC 405862 5-10-5 163 407 426 0 CAAGGCTAGCACCAGCTCCT405863 5-10-5 164 408 427 0 GCAAGGCTAGCACCAGCTCC 405864 5-10-5 165 409428 0 CGCAAGGCTAGCACCAGCTC 405865 5-10-5 166 411 430 0AACGCAAGGCTAGCACCAGC 405866 5-10-5 167 412 431 0 GAACGCAAGGCTAGCACCAG405867 5-10-5 168 413 432 0 GGAACGCAAGGCTAGCACCA 405868 5-10-5 169 414433 0 CGGAACGCAAGGCTAGCACC 405869 5-10-5 218 862 881 0TCGATTTCCCGGTGGTCACT 405870 5-10-5 219 863 882 0 CTCGATTTCCCGGTGGTCAC405871 5-10-5 220 864 883 0 CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 865884 0 CCCTCGATTTCCCGGTGGTC 405873 5-10-5 222 867 886 0TGCCCTCGATTTCCCGGTGG 405874 5-10-5 223 868 887 0 CTGCCCTCGATTTCCCGGTG405875 5-10-5 224 869 888 0 CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 870889 0 CCCTGCCCTCGATTTCCCGG 405877 5-10-5 246 1000 1019 0TTGGCCACGCCGGCATCCCG 405878 5-10-5 247 1001 1020 0 CTTGGCCACGCCGGCATCCC405879 5-10-5 248 1002 1021 0 CCTTGGCCACGCCGGCATCC 405880 5-10-5 2491003 1022 0 CCCTTGGCCACGCCGGCATC 405881 5-10-5 250 1005 1024 0CACCCTTGGCCACGCCGGCA 405882 5-10-5 251 1006 1025 0 GCACCCTTGGCCACGCCGGC405883 5-10-5 252 1007 1026 0 GGCACCCTTGGCCACGCCGG 405884 5-10-5 2531008 1027 0 TGGCACCCTTGGCCACGCCG 405885 5-10-5 346 1560 1579 0GGAACCAGGCCTCATTGATG 405886 5-10-5 347 1561 1580 0 GGGAACCAGGCCTCATTGAT405887 5-10-5 348 1562 1581 0 AGGGAACCAGGCCTCATTGA 405888 5-10-5 3491563 1582 0 CAGGGAACCAGGCCTCATTG 405889 5-10-5 350 1565 1584 0CTCAGGGAACCAGGCCTCAT 405890 5-10-5 351 1566 1585 0 CCTCAGGGAACCAGGCCTCA405891 5-10-5 352 1567 1586 0 TCCTCAGGGAACCAGGCCTC 405892 5-10-5 3531568 1587 0 GTCCTCAGGGAACCAGGCCT 405893 5-10-5 431 2828 2847 0ATGAGGGCCATCAGCACCTT 405894 5-10-5 432 2829 2848 0 GATGAGGGCCATCAGCACCT405895 5-10-5 433 2830 2849 0 AGATGAGGGCCATCAGCACC 405896 5-10-5 4342831 2850 0 GAGATGAGGGCCATCAGCAC 405897 5-10-5 435 2833 2852 0TGGAGATGAGGGCCATCAGC 405898 5-10-5 436 2834 2853 0 CTGGAGATGAGGGCCATCAG405899 5-10-5 437 2835 2854 0 GCTGGAGATGAGGGCCATCA 405900 5-10-5 4382836 2855 0 AGCTGGAGATGAGGGCCATC 405901 5-10-5 439 2902 2921 0GCTAGATGCCATCCAGAAAG 405902 5-10-5 440 2903 2922 0 GGCTAGATGCCATCCAGAAA405903 5-10-5 441 2904 2923 0 TGGCTAGATGCCATCCAGAA 405904 5-10-5 4422905 2924 0 CTGGCTAGATGCCATCCAGA 405905 5-10-5 443 2907 2926 0CTCTGGCTAGATGCCATCCA 405906 5-10-5 444 2908 2927 0 CCTCTGGCTAGATGCCATCC405907 5-10-5 445 2909 2928 0 GCCTCTGGCTAGATGCCATC 405908 5-10-5 4462910 2929 0 AGCCTCTGGCTAGATGCCAT 405909 5-10-5 267 1100 1119 0CAGCTGGCTTTTCCGAATAA 405910 5-10-5 268 1102 1121 0 ACCAGCTGGCTTTTCCGAAT405911 5-10-5 269 1104 1123 0 GGACCAGCTGGCTTTTCCGA 405912 5-10-5 2701108 1127 0 GGCTGGACCAGCTGGCTTTT 405913 5-10-5 275 1212 1231 0CGGTGACCAGCACGACCCCA 405914 5-10-5 276 1214 1233 0 AGCGGTGACCAGCACGACCC405915 5-10-5 277 1216 1235 0 GCAGCGGTGACCAGCACGAC 405916 5-10-5 2781218 1237 0 CGGCAGCGGTGACCAGCACG 405917 5-10-5 280 1222 1241 0TTGCCGGCAGCGGTGACCAG 405918 5-10-5 281 1224 1243 0 AGTTGCCGGCAGCGGTGACC405919 5-10-5 282 1226 1245 0 GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 2831228 1247 0 CGGAAGTTGCCGGCAGCGGT 405921 5-10-5 284 1230 1249 0CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 285 1232 1251 0 GTCCCGGAAGTTGCCGGCAG405923 5-10-5 288 1295 1314 0 GGCATTGGTGGCCCCAACTG 405924 5-10-5 2901318 1337 0 CCCAGGGTCACCGGCTGGTC 405925 5-10-5 292 1322 1341 0AGTCCCCAGGGTCACCGGCT 405926 5-10-5 293 1324 1343 0 AAAGTCCCCAGGGTCACCGG405927 5-10-5 294 1328 1347 0 CCCCAAAGTCCCCAGGGTCA 405928 5-10-5 2951333 1352 0 TTGGTCCCCAAAGTCCCCAG 405929 5-10-5 296 1337 1356 0AAAGTTGGTCCCCAAAGTCC 405930 5-10-5 297 1342 1361 0 CGGCCAAAGTTGGTCCCCAA405931 5-10-5 298 1344 1363 0 AGCGGCCAAAGTTGGTCCCC 405932 5-10-5 2991346 1365 0 ACAGCGGCCAAAGTTGGTCC 405933 5-10-5 300 1348 1367 0ACACAGCGGCCAAAGTTGGT 405934 5-10-5 301 1350 1369 0 CCACACAGCGGCCAAAGTTG405935 5-10-5 302 1354 1373 0 AGGTCCACACAGCGGCCAAA 405936 5-10-5 3041358 1377 0 AAAGAGGTCCACACAGCGGC 405937 5-10-5 306 1387 1406 0GAGGCACCAATGATGTCCTC 405938 5-10-5 307 1391 1410 0 GCTGGAGGCACCAATGATGT405939 5-10-5 308 1393 1412 0 TCGCTGGAGGCACCAATGAT 405940 5-10-5 3091395 1414 0 AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 1397 1416 0GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 1402 1421 0 GTGCTGCAGTCGCTGGAGGC405943 5-10-5 312 1404 1423 0 AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 3141407 1426 0 AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 1409 1428 0AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 1413 1432 0 ACACAAAGCAGGTGCTGCAG405947 5-10-5 317 1415 1434 0 TGACACAAAGCAGGTGCTGC 405948 5-10-5 3191467 1486 0 TCATGGCTGCAATGCCAGCC 405949 5-10-5 320 1472 1491 0CAGCATCATGGCTGCAATGC 405950 5-10-5 321 1474 1493 0 GACAGCATCATGGCTGCAAT405951 5-10-5 322 1476 1495 0 CAGACAGCATCATGGCTGCA 405952 5-10-5 3231480 1499 0 TCGGCAGACAGCATCATGGC 405953 5-10-5 325 1484 1503 0CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 1486 1505 0 TCCGGCTCGGCAGACAGCAT405955 5-10-5 328 1513 1532 0 CTCTGCCTCAACTCGGCCAG 405956 5-10-5 3291515 1534 0 GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 1517 1536 0CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 1519 1538 0 ATCAGTCTCTGCCTCAACTC405959 5-10-5 332 1521 1540 0 GGATCAGTCTCTGCCTCAAC 405960 5-10-5 3331523 1542 0 GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 1525 1544 0AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 1528 1547 0 GAGAAGTGGATCAGTCTCTG405963 5-10-5 337 1532 1551 0 GGCAGAGAAGTGGATCAGTC 405964 5-10-5 3381536 1555 0 CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 1541 1560 0GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 1543 1562 0 ATGACATCTTTGGCAGAGAA405967 5-10-5 341 1547 1566 0 ATTGATGACATCTTTGGCAG 405968 5-10-5 3421549 1568 0 TCATTGATGACATCTTTGGC 405969 5-10-5 343 1554 1573 0AGGCCTCATTGATGACATCT 405970 5-10-5 344 1556 1575 0 CCAGGCCTCATTGATGACAT405971 5-10-5 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG 405972 5-10-5 3571573 1592 0 CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 1578 1597 0GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 1580 1599 0 CAGTACCCGCTGGTCCTCAG405975 5-10-5 361 1642 1661 0 CAAAACAGCTGCCAACCTGC 405976 5-10-5 3621647 1666 0 TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 1649 1668 0AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 1672 1691 0 GGCCCCGAGTGTGCTGACCA405979 5-10-5 366 1677 1696 0 GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 3671679 1698 0 CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 1681 1700 0ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 1685 1704 0 GGCCATCCGTGTAGGCCCCG405983 5-10-5 371 1687 1706 0 GTGGCCATCCGTGTAGGCCC 405984 5-10-5 3721735 1754 0 CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 1737 1756 0AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 1742 1761 0 GGAGAAACTGGAGCAGCTCA405987 5-10-5 375 1744 1763 0 CTGGAGAAACTGGAGCAGCT 405988 5-10-5 3811854 1873 0 GGCAGCACCTGGCAATGGCG 405989 5-10-5 383 1856 1875 0CAGGCAGCACCTGGCAATGG 405990 5-10-5 386 1860 1879 0 GTAGCAGGCAGCACCTGGCA405991 5-10-5 394 1922 1941 0 GACACGGGTCCCCATGCTGG 405992 5-10-5 3961924 1943 0 TGGACACGGGTCCCCATGCT 405993 5-10-5 398 1926 1945 0AGTGGACACGGGTCCCCATG 405994 5-10-5 400 1928 1947 0 GCAGTGGACACGGGTCCCCA405995 5-10-5 402 1930 1949 0 TGGCAGTGGACACGGGTCCC 405996 5-10-5 4122102 2121 0 GACTTTGCATTCCAGACCTG 405997 5-10-5 415 2107 2126 0TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 2313 2332 0 TCCGGCAGCAGATGGCAACG405999 5-10-5 160 298 317 0 GACCGCCTGGAGCTGACGGT 406000 5-10-5 173 482501 0 ATCCTTGGCGCAGCGGTGGA 406001 5-10-5 174 484 503 0GGATCCTTGGCGCAGCGGTG 406002 5-10-5 175 488 507 0 CCACGGATCCTTGGCGCAGC406003 5-10-5 178 555 574 0 CAGTGCGCTCTGACTGCGAG 406004 5-10-5 179 557576 0 GGCAGTGCGCTCTGACTGCG 406005 5-10-5 180 559 578 0CGGGCAGTGCGCTCTGACTG 406006 5-10-5 181 562 581 0 CGGCGGGCAGTGCGCTCTGA406007 5-10-5 183 595 614 0 AGGTATCCCCGGCGGGCAGC 406008 5-10-5 188 602621 0 CTTGGTGAGGTATCCCCGGC 406009 5-10-5 190 604 623 0ATCTTGGTGAGGTATCCCCG 406010 5-10-5 193 609 628 0 GCAGGATCTTGGTGAGGTAT406011 5-10-5 194 611 630 0 ATGCAGGATCTTGGTGAGGT 406012 5-10-5 195 613632 0 ACATGCAGGATCTTGGTGAG 406013 5-10-5 196 617 636 0GAAGACATGCAGGATCTTGG 406014 5-10-5 199 648 667 0 TCTTCACCAGGAAGCCAGGA406015 5-10-5 200 653 672 0 ACTCATCTTCACCAGGAAGC 406016 5-10-5 201 655674 0 CCACTCATCTTCACCAGGAA 406017 5-10-5 203 659 678 0GTCGCCACTCATCTTCACCA 406018 5-10-5 204 661 680 0 AGGTCGCCACTCATCTTCAC406019 5-10-5 205 663 682 0 GCAGGTCGCCACTCATCTTC 406020 5-10-5 206 665684 0 CAGCAGGTCGCCACTCATCT 406021 5-10-5 210 782 801 0TTCATCCGCCCGGTACCGTG 406022 5-10-5 211 784 803 0 TATTCATCCGCCCGGTACCG406023 5-10-5 212 787 806 0 TGGTATTCATCCGCCCGGTA 406024 5-10-5 213 789808 0 GCTGGTATTCATCCGCCCGG 406025 5-10-5 216 832 851 0GTGTCTAGGAGATACACCTC 406026 5-10-5 217 837 856 0 TGCTGGTGTCTAGGAGATAC406027 5-10-5 226 874 893 0 ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 876895 0 CCATGACCCTGCCCTCGATT 406029 5-10-5 228 878 897 0GACCATGACCCTGCCCTCGA 406030 5-10-5 229 882 901 0 CGGTGACCATGACCCTGCCC406031 5-10-5 230 884 903 0 GTCGGTGACCATGACCCTGC 406032 5-10-5 232 888907 0 CGAAGTCGGTGACCATGACC 406033 5-10-5 237 967 986 0CCTGCCAGGTGGGTGCCATG 406034 5-10-5 238 972 991 0 CCACCCCTGCCAGGTGGGTG406035 5-10-5 239 977 996 0 GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 9891008 0 GGCATCCCGGCCGCTGACCA 406037 5-10-5 242 992 1011 0GCCGGCATCCCGGCCGCTGA 406038 5-10-5 245 999 1018 0 TGGCCACGCCGGCATCCCGG406039 5-10-5 257 1036 1055 0 CAGTTGAGCACGCGCAGGCT 406040 5-10-5 2581038 1057 0 GGCAGTTGAGCACGCGCAGG 406041 5-10-5 259 1042 1061 0CCTTGGCAGTTGAGCACGCG 406042 5-10-5 260 1047 1066 0 CCTTCCCTTGGCAGTTGAGC406043 5-10-5 261 1051 1070 0 GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 2621053 1072 0 CCGTGCCCTTCCCTTGGCAG 406045 5-10-5 266 1096 1115 0TGGCTTTTCCGAATAAACTC 408642 5-10-5 264 1076 1095 0 CAGGCCTATGAGGGTGCCGC408653 5-10-5 354 1570 1589 0 TGGTCCTCAGGGAACCAGGC 409126 5-10-5 4471004 1023 1 ACCCTTGGTCACGCCGGCAT 410529 5-10-5 184 597 616 0TGAGGTATCCCCGGCGGGCA 410530 5-10-5 185 598 617 0 GTGAGGTATCCCCGGCGGGC410531 5-10-5 186 599 618 0 GGTGAGGTATCCCCGGCGGG 410532 5-10-5 187 601620 0 TTGGTGAGGTATCCCCGGCG 410533 5-10-5 189 603 622 0TCTTGGTGAGGTATCCCCGG 410534 5-10-5 191 605 624 0 GATCTTGGTGAGGTATCCCC410535 5-10-5 192 607 626 0 AGGATCTTGGTGAGGTATCC 410536 5-10-5 243 9971016 0 GCCACGCCGGCATCCCGGCC 410537 5-10-5 244 998 1017 0GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 1009 1028 0 CTGGCACCCTTGGCCACGCC410539 5-10-5 255 1010 1029 0 GCTGGCACCCTTGGCCACGC 410540 5-10-5 3561572 1591 0 GCTGGTCCTCAGGGAACCAG 410541 5-10-5 376 1849 1868 0CACCTGGCAATGGCGTAGAC 410542 5-10-5 377 1850 1869 0 GCACCTGGCAATGGCGTAGA410543 5-10-5 378 1851 1870 0 AGCACCTGGCAATGGCGTAG 410544 5-10-5 3791852 1871 0 CAGCACCTGGCAATGGCGTA 410545 5-10-5 380 1853 1872 0GCAGCACCTGGCAATGGCGT 410546 5-10-5 382 1855 1874 0 AGGCAGCACCTGGCAATGGC410547 5-10-5 384 1857 1876 0 GCAGGCAGCACCTGGCAATG 410548 5-10-5 3851859 1878 0 TAGCAGGCAGCACCTGGCAA 410549 5-10-5 388 1915 1934 0GTCCCCATGCTGGCCTCAGC 410550 5-10-5 389 1916 1935 0 GGTCCCCATGCTGGCCTCAG410551 5-10-5 390 1917 1936 0 GGGTCCCCATGCTGGCCTCA 410552 5-10-5 3911918 1937 0 CGGGTCCCCATGCTGGCCTC 410553 5-10-5 392 1919 1938 0ACGGGTCCCCATGCTGGCCT 410554 5-10-5 393 1921 1940 0 ACACGGGTCCCCATGCTGGC410555 5-10-5 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410556 5-10-5 3971925 1944 0 GTGGACACGGGTCCCCATGC 410557 5-10-5 399 1927 1946 0CAGTGGACACGGGTCCCCAT 410558 5-10-5 401 1929 1948 0 GGCAGTGGACACGGGTCCCC410559 5-10-5 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410560 5-10-5 4041932 1951 0 GGTGGCAGTGGACACGGGTC 410561 5-10-5 405 1933 1952 0TGGTGGCAGTGGACACGGGT 410562 5-10-5 411 2101 2120 0 ACTTTGCATTCCAGACCTGG410563 5-10-5 413 2103 2122 0 TGACTTTGCATTCCAGACCT 410564 5-10-5 4142104 2123 0 TTGACTTTGCATTCCAGACC 410565 5-10-5 419 2305 2324 0CAGATGGCAACGGCTGTCAC 410566 5-10-5 420 2306 2325 0 GCAGATGGCAACGGCTGTCA410567 5-10-5 421 2307 2326 0 AGCAGATGGCAACGGCTGTC 410568 5-10-5 4222308 2327 0 CAGCAGATGGCAACGGCTGT 410569 5-10-5 423 2309 2328 0GCAGCAGATGGCAACGGCTG 410570 5-10-5 424 2311 2330 0 CGGCAGCAGATGGCAACGGC410571 5-10-5 425 2312 2331 0 CCGGCAGCAGATGGCAACGG 410572 5-10-5 4272314 2333 0 CTCCGGCAGCAGATGGCAAC 410573 5-10-5 428 2315 2334 0GCTCCGGCAGCAGATGGCAA 410730 5-10-5 202 657 676 0 CGCCACTCATCTTCACCAGG410731 5-10-5 207 667 686 0 TCCAGCAGGTCGCCACTCAT 410732 5-10-5 214 791810 0 GGGCTGGTATTCATCCGCCC 410733 5-10-5 231 886 905 0AAGTCGGTGACCATGACCCT 410734 5-10-5 279 1219 1238 0 CCGGCAGCGGTGACCAGCAC410735 5-10-5 291 1320 1339 0 TCCCCAGGGTCACCGGCTGG 410736 5-10-5 3031356 1375 0 AGAGGTCCACACAGCGGCCA 410737 5-10-5 313 1406 1425 0GCAGGTGCTGCAGTCGCTGG 410738 5-10-5 324 1482 1501 0 GCTCGGCAGACAGCATCATG410739 5-10-5 336 1530 1549 0 CAGAGAAGTGGATCAGTCTC 410740 5-10-5 3451558 1577 0 AACCAGGCCTCATTGATGAC 410741 5-10-5 369 1683 1702 0CCATCCGTGTAGGCCCCGAG 410742 5-10-5 159 294 313 0 GCCTGGAGCTGACGGTGCCC410743 5-10-5 170 421 440 0 TCCTCCTCGGAACGCAAGGC 410744 5-10-5 171 446465 0 GTGCTCGGGTGCTTCGGCCA 410745 5-10-5 172 466 485 0TGGAAGGTGGCTGTGGTTCC 410746 5-10-5 176 507 526 0 CGTAGGTGCCAGGCAACCTC410747 5-10-5 177 545 564 0 TGACTGCGAGAGGTGGGTCT 410748 5-10-5 182 591610 0 ATCCCCGGCGGGCAGCCTGG 410749 5-10-5 197 628 647 0AGAAGGCCATGGAAGACATG 410750 5-10-5 198 638 657 0 GAAGCCAGGAAGAAGGCCAT410751 5-10-5 208 685 704 0 GGCAACTTCAAGGCCAGCTC 410752 5-10-5 209 724743 0 GCAAAGACAGAGGAGTCCTC 410753 5-10-5 215 821 840 0ATACACCTCCACCAGGCTGC 410754 5-10-5 233 898 917 0 GGCACATTCTCGAAGTCGGT410755 5-10-5 234 933 952 0 TGGCCTGTCTGTGGAAGCGG 410756 5-10-5 235 960979 0 GGTGGGTGCCATGACTGTCA 410757 5-10-5 240 985 1004 0TCCCGGCCGCTGACCACCCC 410758 5-10-5 256 1015 1034 0 CGCATGCTGGCACCCTTGGC410759 5-10-5 263 1064 1083 0 GGTGCCGCTAACCGTGCCCT 410760 5-10-5 2651088 1107 0 CCGAATAAACTCCAGGCCTA 410761 5-10-5 271 1119 1138 0GTGGCCCCACAGGCTGGACC 410762 5-10-5 272 1132 1151 0 AGCAGCACCACCAGTGGCCC410763 5-10-5 273 1154 1173 0 GCTGTACCCACCCGCCAGGG 410764 5-10-5 2741200 1219 0 CGACCCCAGCCCTCGCCAGG 410765 5-10-5 286 1273 1292 0ATGACCTCGGGAGCTGAGGC 410766 5-10-5 287 1283 1302 0 CCCAACTGTGATGACCTCGG410767 5-10-5 289 1305 1324 0 GCTGGTCTTGGGCATTGGTG 410768 5-10-5 3051380 1399 0 CAATGATGTCCTCCCCTGGG 410769 5-10-5 318 1425 1444 0TCCCACTCTGTGACACAAAG 410770 5-10-5 327 1500 1519 0 CGGCCAGGGTGAGCTCCGGC410771 5-10-5 360 1628 1647 0 ACCTGCCCCATGGGTGCTGG 410772 5-10-5 3641660 1679 0 GCTGACCATACAGTCCTGCA 410773 5-10-5 387 1905 1924 0TGGCCTCAGCTGGTGGAGCT 410774 5-10-5 406 1936 1955 0 TGTTGGTGGCAGTGGACACG410775 5-10-5 407 1962 1981 0 AGCTGCAGCCTGTGAGGACG 410776 5-10-5 4081990 2009 0 GTGCCAAGGTCCTCCACCTC 410777 5-10-5 409 2010 2029 0TCAGCACAGGCGGCTTGTGG 410778 5-10-5 410 2040 2059 0 CCACGCACTGGTTGGGCTGA410779 5-10-5 416 2120 2139 0 CGGGATTCCATGCTCCTTGA 410780 5-10-5 4172150 2169 0 GCAGGCCACGGTCACCTGCT 410781 5-10-5 418 2187 2206 0GGAGGGCACTGCAGCCAGTC 410782 5-10-5 429 2325 2344 0 CCAGGTGCCGGCTCCGGCAG410783 5-10-5 430 2335 2354 0 GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 1. In certain such embodiments, thenucleotide sequences illustrated in Table 2 have a 3-14-3 gap-widenedmotif. Table 4 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 1, having a 3-14-3 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 4 Gapmer antisense compounds having a 3-14-3 motif targeted to SEQID NO: 1 5′ Target Site 3′ Target Site SEQ to SEQ ID to SEQ ID Isis NoMotif ID NO NO: 1 NO: 1 Mismatches Sequence (5′-3′) 399871 3-14-3 4 135154 0 GCGCGGAATCCTGGCTGGGA 399872 3-14-3 5 242 261 0GAGGAGACCTAGAGGCCGTG 399873 3-14-3 6 300 319 0 AGGACCGCCTGGAGCTGACG399874 3-14-3 7 410 429 0 ACGCAAGGCTAGCACCAGCT 399949 3-14-3 8 417 436 0CCTCGGAACGCAAGGCTAGC 399875 3-14-3 9 480 499 0 CCTTGGCGCAGCGGTGGAAG399876 3-14-3 10 561 580 0 GGCGGGCAGTGCGCTCTGAC 399877 3-14-3 11 600 6190 TGGTGAGGTATCCCCGGCGG 399950 3-14-3 12 606 625 0 GGATCTTGGTGAGGTATCCC399951 3-14-3 13 615 634 0 AGACATGCAGGATCTTGGTG 399878 3-14-3 14 620 6390 ATGGAAGACATGCAGGATCT 399879 3-14-3 15 646 665 0 TTCACCAGGAAGCCAGGAAG399952 3-14-3 16 651 670 0 TCATCTTCACCAGGAAGCCA 399880 3-14-3 17 705 7240 CCTCGATGTAGTCGACATGG 399881 3-14-3 18 785 804 0 GTATTCATCCGCCCGGTACC399882 3-14-3 19 835 854 0 CTGGTGTCTAGGAGATACAC 399953 3-14-3 20 840 8590 GTATGCTGGTGTCTAGGAGA 399883 3-14-3 21 860 879 0 GATTTCCCGGTGGTCACTCT399954 3-14-3 22 866 885 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 23 880 8990 GTGACCATGACCCTGCCCTC 399884 3-14-3 24 890 909 0 CTCGAAGTCGGTGACCATGA399885 3-14-3 25 923 942 0 GTGGAAGCGGGTCCCGTCCT 399886 3-14-3 26 970 9890 ACCCCTGCCAGGTGGGTGCC 399956 3-14-3 27 975 994 0 TGACCACCCCTGCCAGGTGG399887 3-14-3 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 10401059 0 TTGGCAGTTGAGCACGCGCA 399957 3-14-3 30 1045 1064 0TTCCCTTGGCAGTTGAGCAC 399889 3-14-3 31 1077 1096 0 CCAGGCCTATGAGGGTGCCG399890 3-14-3 32 1098 1117 0 GCTGGCTTTTCCGAATAAAC 399891 3-14-3 33 12101229 0 GTGACCAGCACGACCCCAGC 399892 3-14-3 149 1297 1316 0TGGGCATTGGTGGCCCCAAC 399893 3-14-3 34 1326 1345 0 CCAAAGTCCCCAGGGTCACC399894 3-14-3 128 1330 1349 0 GTCCCCAAAGTCCCCAGGGT 399958 3-14-3 35 13351354 0 AGTTGGTCCCCAAAGTCCCC 399895 3-14-3 36 1340 1359 0GCCAAAGTTGGTCCCCAAAG 399959 3-14-3 37 1352 1371 0 GTCCACACAGCGGCCAAAGT399896 3-14-3 38 1361 1380 0 GGCAAAGAGGTCCACACAGC 399897 3-14-3 39 13891408 0 TGGAGGCACCAATGATGTCC 399960 3-14-3 40 1400 1419 0GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 1411 1430 0 ACAAAGCAGGTGCTGCAGTC399898 3-14-3 101 1465 1484 0 ATGGCTGCAATGCCAGCCAC 399962 3-14-3 42 14701489 0 GCATCATGGCTGCAATGCCA 399963 3-14-3 43 1478 1497 0GGCAGACAGCATCATGGCTG 399964 3-14-3 44 1526 1545 0 GAAGTGGATCAGTCTCTGCC399899 3-14-3 45 1534 1553 0 TTGGCAGAGAAGTGGATCAG 399965 3-14-3 46 15391558 0 CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 1545 1564 0TGATGACATCTTTGGCAGAG 399967 3-14-3 48 1552 1571 0 GCCTCATTGATGACATCTTT399968 3-14-3 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 399900 3-14-3 50 15691588 0 GGTCCTCAGGGAACCAGGCC 399901 3-14-3 87 1576 1595 0ACCCGCTGGTCCTCAGGGAA 399969 3-14-3 51 1583 1602 0 GGTCAGTACCCGCTGGTCCT399902 3-14-3 119 1605 1624 0 GCAGGGCGGCCACCAGGTTG 399903 3-14-3 52 16401659 0 AAACAGCTGCCAACCTGCCC 399970 3-14-3 53 1645 1664 0CTGCAAAACAGCTGCCAACC 399904 3-14-3 54 1675 1694 0 GTAGGCCCCGAGTGTGCTGA399971 3-14-3 55 1740 1759 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 56 17461765 0 TCCTGGAGAAACTGGAGCAG 399905 3-14-3 57 1812 1831 0CGTTGTGGGCCCGGCAGACC 399906 3-14-3 58 1858 1877 0 AGCAGGCAGCACCTGGCAAT399907 3-14-3 59 1920 1939 0 CACGGGTCCCCATGCTGGCC 399908 3-14-3 60 21002119 0 CTTTGCATTCCAGACCTGGG 399973 3-14-3 61 2105 2124 0CTTGACTTTGCATTCCAGAC 399909 3-14-3 62 2310 2329 0 GGCAGCAGATGGCAACGGCT399910 3-14-3 154 2410 2429 0 TTTTAAAGCTCAGCCCCAGC 399974 3-14-3 63 24152434 0 AACCATTTTAAAGCTCAGCC 399911 3-14-3 64 2504 2523 0TCAAGGGCCAGGCCAGCAGC 399975 3-14-3 65 2509 2528 0 CCCACTCAAGGGCCAGGCCA399976 3-14-3 122 2582 2601 0 GGAGGGAGCTTCCTGGCACC 399912 3-14-3 66 25972616 0 ATGCCCCACAGTGAGGGAGG 399913 3-14-3 67 2606 2625 0AATGGTGAAATGCCCCACAG 399914 3-14-3 153 2649 2668 0 TTGGGAGCAGCTGGCAGCAC399915 3-14-3 68 2750 2769 0 CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 28322851 0 GGAGATGAGGGCCATCAGCA 399917 3-14-3 70 2900 2919 0TAGATGCCATCCAGAAAGCT 399977 3-14-3 71 2906 2925 0 TCTGGCTAGATGCCATCCAG399918 3-14-3 72 2983 3002 0 GGCATAGAGCAGAGTAAAGG 399978 3-14-3 73 29883007 0 AGCCTGGCATAGAGCAGAGT 399979 3-14-3 135 2994 3013 0TAGCACAGCCTGGCATAGAG 399919 3-14-3 112 3227 3246 0 GAAGAGGCTTGGCTTCAGAG399980 3-14-3 74 3233 3252 0 AAGTAAGAAGAGGCTTGGCT 399920 3-14-3 75 34373456 0 GCTCAAGGAGGGACAGTTGT 399921 3-14-3 76 3472 3491 0AAAGATAAATGTCTGCTTGC 399981 3-14-3 77 3477 3496 0 ACCCAAAAGATAAATGTCTG399922 3-14-3 78 3543 3562 0 TCTTCAAGTTACAAAAGCAA 399982 3-14-3 99 35503569 0 ATAAATATCTTCAAGTTACA 410574 3-14-3 184 597 616 0TGAGGTATCCCCGGCGGGCA 410575 3-14-3 185 598 617 0 GTGAGGTATCCCCGGCGGGC410576 3-14-3 186 599 618 0 GGTGAGGTATCCCCGGCGGG 410577 3-14-3 187 601620 0 TTGGTGAGGTATCCCCGGCG 410578 3-14-3 188 602 621 0CTTGGTGAGGTATCCCCGGC 410579 3-14-3 189 603 622 0 TCTTGGTGAGGTATCCCCGG410580 3-14-3 190 604 623 0 ATCTTGGTGAGGTATCCCCG 410581 3-14-3 191 605624 0 GATCTTGGTGAGGTATCCCC 410582 3-14-3 192 607 626 0AGGATCTTGGTGAGGTATCC 405604 3-14-3 236 963 982 0 CCAGGTGGGTGCCATGACTG410583 3-14-3 243 997 1016 0 GCCACGCCGGCATCCCGGCC 410584 3-14-3 244 9981017 0 GGCCACGCCGGCATCCCGGC 410585 3-14-3 245 999 1018 0TGGCCACGCCGGCATCCCGG 410586 3-14-3 246 1000 1019 0 TTGGCCACGCCGGCATCCCG410587 3-14-3 247 1001 1020 0 CTTGGCCACGCCGGCATCCC 410588 3-14-3 2481002 1021 0 CCTTGGCCACGCCGGCATCC 410589 3-14-3 249 1003 1022 0CCCTTGGCCACGCCGGCATC 410590 3-14-3 250 1005 1024 0 CACCCTTGGCCACGCCGGCA410591 3-14-3 251 1006 1025 0 GCACCCTTGGCCACGCCGGC 410592 3-14-3 2521007 1026 0 GGCACCCTTGGCCACGCCGG 410593 3-14-3 253 1008 1027 0TGGCACCCTTGGCCACGCCG 410594 3-14-3 254 1009 1028 0 CTGGCACCCTTGGCCACGCC410595 3-14-3 255 1010 1029 0 GCTGGCACCCTTGGCCACGC 410596 3-14-3 3481562 1581 0 AGGGAACCAGGCCTCATTGA 410597 3-14-3 349 1563 1582 0CAGGGAACCAGGCCTCATTG 410598 3-14-3 350 1565 1584 0 CTCAGGGAACCAGGCCTCAT410599 3-14-3 351 1566 1585 0 CCTCAGGGAACCAGGCCTCA 410600 3-14-3 3521567 1586 0 TCCTCAGGGAACCAGGCCTC 410601 3-14-3 353 1568 1587 0GTCCTCAGGGAACCAGGCCT 410602 3-14-3 354 1570 1589 0 TGGTCCTCAGGGAACCAGGC410603 3-14-3 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG 410604 3-14-3 3561572 1591 0 GCTGGTCCTCAGGGAACCAG 405641 3-14-3 373 1737 1756 0AACTGGAGCAGCTCAGCAGC 410605 3-14-3 376 1849 1868 0 CACCTGGCAATGGCGTAGAC410606 3-14-3 377 1850 1869 0 GCACCTGGCAATGGCGTAGA 410607 3-14-3 3781851 1870 0 AGCACCTGGCAATGGCGTAG 410608 3-14-3 379 1852 1871 0CAGCACCTGGCAATGGCGTA 410609 3-14-3 380 1853 1872 0 GCAGCACCTGGCAATGGCGT410610 3-14-3 381 1854 1873 0 GGCAGCACCTGGCAATGGCG 410611 3-14-3 3821855 1874 0 AGGCAGCACCTGGCAATGGC 410612 3-14-3 383 1856 1875 0CAGGCAGCACCTGGCAATGG 410613 3-14-3 384 1857 1876 0 GCAGGCAGCACCTGGCAATG410614 3-14-3 385 1859 1878 0 TAGCAGGCAGCACCTGGCAA 410615 3-14-3 3881915 1934 0 GTCCCCATGCTGGCCTCAGC 410616 3-14-3 389 1916 1935 0GGTCCCCATGCTGGCCTCAG 410617 3-14-3 390 1917 1936 0 GGGTCCCCATGCTGGCCTCA410618 3-14-3 391 1918 1937 0 CGGGTCCCCATGCTGGCCTC 410619 3-14-3 3921919 1938 0 ACGGGTCCCCATGCTGGCCT 410620 3-14-3 393 1921 1940 0ACACGGGTCCCCATGCTGGC 410621 3-14-3 394 1922 1941 0 GACACGGGTCCCCATGCTGG410622 3-14-3 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410623 3-14-3 3961924 1943 0 TGGACACGGGTCCCCATGCT 410624 3-14-3 397 1925 1944 0GTGGACACGGGTCCCCATGC 410625 3-14-3 398 1926 1945 0 AGTGGACACGGGTCCCCATG410626 3-14-3 399 1927 1946 0 CAGTGGACACGGGTCCCCAT 410627 3-14-3 4001928 1947 0 GCAGTGGACACGGGTCCCCA 410628 3-14-3 401 1929 1948 0GGCAGTGGACACGGGTCCCC 410629 3-14-3 402 1930 1949 0 TGGCAGTGGACACGGGTCCC410630 3-14-3 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410631 3-14-3 4041932 1951 0 GGTGGCAGTGGACACGGGTC 410632 3-14-3 405 1933 1952 0TGGTGGCAGTGGACACGGGT 410633 3-14-3 411 2101 2120 0 ACTTTGCATTCCAGACCTGG410634 3-14-3 412 2102 2121 0 GACTTTGCATTCCAGACCTG 410635 3-14-3 4132103 2122 0 TGACTTTGCATTCCAGACCT 410636 3-14-3 414 2104 2123 0TTGACTTTGCATTCCAGACC 410637 3-14-3 419 2305 2324 0 CAGATGGCAACGGCTGTCAC410638 3-14-3 420 2306 2325 0 GCAGATGGCAACGGCTGTCA 410639 3-14-3 4212307 2326 0 AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 2308 2327 0CAGCAGATGGCAACGGCTGT 410641 3-14-3 423 2309 2328 0 GCAGCAGATGGCAACGGCTG410642 3-14-3 424 2311 2330 0 CGGCAGCAGATGGCAACGGC 410643 3-14-3 4252312 2331 0 CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 2313 2332 0TCCGGCAGCAGATGGCAACG 410645 3-14-3 427 2314 2333 0 CTCCGGCAGCAGATGGCAAC410646 3-14-3 428 2315 2334 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 1. In certain such embodiments, thenucleotide sequences illustrated in Table 2 have a 2-13-5 gap-widenedmotif. Table 5 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 1, having a 2-13-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 5 Gapmer antisense compounds having a 2-13-5 motif targeted to SEQID NO: 1 5′ start site SEQ to SEQ ID 3′ Start Site to ISIS No. Motif IDNO NO: 1 SEQ ID NO: 1 Mismatches Sequences (5′ to 3′) 410647 2-13-5 184597 616 0 TGAGGTATCCCCGGCGGGCA 410648 2-13-5 185 598 617 0GTGAGGTATCCCCGGCGGGC 410649 2-13-5 186 599 618 0 GGTGAGGTATCCCCGGCGGG410650 2-13-5 11 600 619 0 TGGTGAGGTATCCCCGGCGG 410651 2-13-5 187 601620 0 TTGGTGAGGTATCCCCGGCG 410652 2-13-5 188 602 621 0CTTGGTGAGGTATCCCCGGC 410653 2-13-5 189 603 622 0 TCTTGGTGAGGTATCCCCGG410654 2-13-5 190 604 623 0 ATCTTGGTGAGGTATCCCCG 410655 2-13-5 191 605624 0 GATCTTGGTGAGGTATCCCC 410656 2-13-5 12 606 625 0GGATCTTGGTGAGGTATCCC 410657 2-13-5 192 607 626 0 AGGATCTTGGTGAGGTATCC410658 2-13-5 243 997 1016 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 244 9981017 0 GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 999 1018 0TGGCCACGCCGGCATCCCGG 410661 2-13-5 246 1000 1019 0 TTGGCCACGCCGGCATCCCG410662 2-13-5 247 1001 1020 0 CTTGGCCACGCCGGCATCCC 410663 2-13-5 2481002 1021 0 CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 1003 1022 0CCCTTGGCCACGCCGGCATC 410665 2-13-5 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT410666 2-13-5 250 1005 1024 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 2511006 1025 0 GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 1007 1026 0GGCACCCTTGGCCACGCCGG 410669 2-13-5 253 1008 1027 0 TGGCACCCTTGGCCACGCCG410670 2-13-5 254 1009 1028 0 CTGGCACCCTTGGCCACGCC 410671 2-13-5 2551010 1029 0 GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 1562 1581 0AGGGAACCAGGCCTCATTGA 410673 2-13-5 349 1563 1582 0 CAGGGAACCAGGCCTCATTG410674 2-13-5 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 410675 2-13-5 350 15651584 0 CTCAGGGAACCAGGCCTCAT 410676 2-13-5 351 1566 1585 0CCTCAGGGAACCAGGCCTCA 410677 2-13-5 352 1567 1586 0 TCCTCAGGGAACCAGGCCTC410678 2-13-5 353 1568 1587 0 GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 15691588 0 GGTCCTCAGGGAACCAGGCC 410680 2-13-5 354 1570 1589 0TGGTCCTCAGGGAACCAGGC 410681 2-13-5 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG410682 2-13-5 356 1572 1591 0 GCTGGTCCTCAGGGAACCAG 410683 2-13-5 3761849 1868 0 CACCTGGCAATGGCGTAGAC 410684 2-13-5 377 1850 1869 0GCACCTGGCAATGGCGTAGA 410685 2-13-5 378 1851 1870 0 AGCACCTGGCAATGGCGTAG410686 2-13-5 379 1852 1871 0 CAGCACCTGGCAATGGCGTA 410687 2-13-5 3801853 1872 0 GCAGCACCTGGCAATGGCGT 410688 2-13-5 381 1854 1873 0GGCAGCACCTGGCAATGGCG 410689 2-13-5 382 1855 1874 0 AGGCAGCACCTGGCAATGGC410690 2-13-5 383 1856 1875 0 CAGGCAGCACCTGGCAATGG 410691 2-13-5 3841857 1876 0 GCAGGCAGCACCTGGCAATG 410692 2-13-5 58 1858 1877 0AGCAGGCAGCACCTGGCAAT 410693 2-13-5 385 1859 1878 0 TAGCAGGCAGCACCTGGCAA410694 2-13-5 388 1915 1934 0 GTCCCCATGCTGGCCTCAGC 410695 2-13-5 3891916 1935 0 GGTCCCCATGCTGGCCTCAG 410696 2-13-5 390 1917 1936 0GGGTCCCCATGCTGGCCTCA 410697 2-13-5 391 1918 1937 0 CGGGTCCCCATGCTGGCCTC410698 2-13-5 392 1919 1938 0 ACGGGTCCCCATGCTGGCCT 410699 2-13-5 59 19201939 0 CACGGGTCCCCATGCTGGCC 410700 2-13-5 393 1921 1940 0ACACGGGTCCCCATGCTGGC 410701 2-13-5 394 1922 1941 0 GACACGGGTCCCCATGCTGG410702 2-13-5 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410703 2-13-5 3961924 1943 0 TGGACACGGGTCCCCATGCT 410704 2-13-5 397 1925 1944 0GTGGACACGGGTCCCCATGC 410705 2-13-5 398 1926 1945 0 AGTGGACACGGGTCCCCATG410706 2-13-5 399 1927 1946 0 CAGTGGACACGGGTCCCCAT 410707 2-13-5 4001928 1947 0 GCAGTGGACACGGGTCCCCA 410708 2-13-5 401 1929 1948 0GGCAGTGGACACGGGTCCCC 410709 2-13-5 402 1930 1949 0 TGGCAGTGGACACGGGTCCC410710 2-13-5 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410711 2-13-5 4041932 1951 0 GGTGGCAGTGGACACGGGTC 410712 2-13-5 405 1933 1952 0TGGTGGCAGTGGACACGGGT 410713 2-13-5 60 2100 2119 0 CTTTGCATTCCAGACCTGGG410714 2-13-5 411 2101 2120 0 ACTTTGCATTCCAGACCTGG 410715 2-13-5 4122102 2121 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 413 2103 2122 0TGACTTTGCATTCCAGACCT 410717 2-13-5 414 2104 2123 0 TTGACTTTGCATTCCAGACC410718 2-13-5 61 2105 2124 0 CTTGACTTTGCATTCCAGAC 410719 2-13-5 419 23052324 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 420 2306 2325 0GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 2307 2326 0 AGCAGATGGCAACGGCTGTC410722 2-13-5 422 2308 2327 0 CAGCAGATGGCAACGGCTGT 410723 2-13-5 4232309 2328 0 GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 2310 2329 0GGCAGCAGATGGCAACGGCT 410725 2-13-5 424 2311 2330 0 CGGCAGCAGATGGCAACGGC410726 2-13-5 425 2312 2331 0 CCGGCAGCAGATGGCAACGG 410727 2-13-5 4262313 2332 0 TCCGGCAGCAGATGGCAACG 410728 2-13-5 427 2314 2333 0CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 2315 2334 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 1. In certain such embodiments, thenucleotide sequences illustrated in Table 2 have a 3-13-4 gap-widenedmotif. Table 6 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 1, having a 3-13-4 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 6 Gapmer antisense compounds having a 3-13-4 motif targeted to SEQID NO: 1 SEQ ID 5′ start 3′ Start Oligo Motif NO: site Site MismatchesOligoSeq 405526 3-13-4 236 963 982 0 CCAGGTGGGTGCCATGACTG 405557 3-13-450 1569 1588 0 GGTCCTCAGGGAACCAGGCC 405564 3-13-4 373 1737 1756 0AACTGGAGCAGCTCAGCAGC

The following embodiments set forth target regions of PCSK9 nucleicacids. Also illustrated are examples of antisense compounds targeted tothe target regions. It is understood that the sequence set forth in eachSEQ ID NO is independent of any modification to a sugar moiety, aninternucleoside linkage, or a nucleobase. As such, antisense compoundsdefined by a SEQ ID NO may comprise, independently, one or moremodifications to a sugar moiety, an internucleoside linkage, or anucleobase. Antisense compounds described by Isis Number (Isis No)indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526,410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622,600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647,638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917,835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909,923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064,970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024,997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021,1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034,1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251,1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345,1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416,1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519,1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568,1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592,1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666,1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876,1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879,1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945,1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950,1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059,2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332,2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334,2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855,2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013,3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, a target region is nucleotides 294-317 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 294-317 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 159 or 160. In certainsuch embodiments, an antisense compound targeted to nucleotides 294-317of SEQ ID NO: 1 is selected from ISIS NOs: 410742 or 405999.

In certain embodiments, a target region is nucleotides 406-440 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 406-440 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 7, 8, 162, 163, 164, 165,166, 167, 168, 169 or 170. In certain such embodiments, an antisensecompound targeted to nucleotides 406-440 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405861, 405862, 405863, 405864, 395152, 399874, 405865,405866, 405867, 405868, 399793, 399949, or 410743.

In certain embodiments, a target region is nucleotides 406-526 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 406-526 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 162, 163, 164,165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, or 176. Incertain such embodiments, an antisense compound targeted to nucleotides406-526 of SEQ ID NO: 1 is selected from ISIS NOs: 405861, 405862,405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793,399950, 410743, 410744, 410745, 395153, 399875, 406000, 406001, 406002or 410746.

In certain embodiments, a target region is nucleotides 410-436 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 410-436 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168, 169or 169. In certain such embodiments, an antisense compound targeted tonucleotides 410-436 of SEQ ID NO: 1 is selected from ISIS NOs: 395152,399874, 405865, 405866, 405867, 405868, or 399793.

In certain embodiments, a target region is nucleotides 410-499 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 410-499 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 166, 167, 169,170, 171 or 172. In certain such embodiments, an antisense compoundtargeted to nucleotides 410-499 of SEQ ID NO: 1 is selected from ISISNOs: 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399949,410743, 410744, 410745, 395153, or 399875.

In certain embodiments, a target region is nucleotides 446-526 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 446-526 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 9, 171, 172, 173, 174, 175or 176. In certain such embodiments, an antisense compound targeted tonucleotides 446-526 of SEQ ID NO: 1 is selected from ISIS NOs: 410744,410745, 395153, 399875, 406000, 406001, 406002, or 410746.

In certain embodiments, a target region is nucleotides 545-581 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 545-581 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or181. In certain such embodiments, an antisense compound targeted tonucleotides 545-581 of SEQ ID NO: 1 is selected from ISIS NOs: 410747,406003, 406004, 406005, 395154, 399876, or 406006.

In certain embodiments, a target region is nucleotides 591-619 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 591-619 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 182, 183, 184, 185 or186. In certain such embodiments, an antisense compound targeted tonucleotides 591-619 of SEQ ID NO: 1 is selected from ISIS NOs: 410748,406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576,410649, 395155, 399877, or 410650.

In certain embodiments, a target region is nucleotides 591-704 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 591-704 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16,182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195,196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, or 208. Incertain such embodiments, an antisense compound targeted to nucleotides591-704 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007,410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649,395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652,410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655,399794, 399950, 410656, or 410751.

In certain embodiments, a target region is nucleotides 591-743 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 591-743 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16,17, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or209. In certain such embodiments, an antisense compound targeted tonucleotides 591-743 of SEQ ID NO: 1 is selected from ISIS NOs: 410748,406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576,410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578,410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581,410655, 399794, 399950, 410656, or 410752.

In certain embodiments, a target region is nucleotides 595-622 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 595-622 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 183, 184, 185, 186,187, 188, or 189. In certain such embodiments, an antisense compoundtargeted to nucleotides 595-622 of SEQ ID NO: 1 is selected from ISISNOs: 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531,410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008,410578, 410652, 410533, 410579, or 410653.

In certain embodiments, a target region is nucleotides 600-626 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 600-626 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 87, 188, 189, 190,191, or 192. In certain such embodiments, an antisense compound targetedto nucleotides 600-626 of SEQ ID NO: 1 is selected from ISIS NOs:395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652,410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655,399794, 399950, 410656, 410535, 410582, or 410657.

In certain embodiments, a target region is nucleotides 600-639 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 600-639 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 187, 188,189, 190, 191, 192, 193, 194, 195, or 196. In certain such embodiments,an antisense compound targeted to nucleotides 600-639 of SEQ ID NO: 1 isselected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651,406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654,410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657,406010, 406011, 406012, 399795, 399951, 406013, 395156, or 399878.

In certain embodiments, a target region is nucleotides 600-670 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 600-670 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16,187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199. Incertain such embodiments, an antisense compound targeted to nucleotides600-670 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877,410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579,410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950,410656, 410535, 410582, 410657, 406010, 406011, 406012, 399795, 399951,406013, 395156, 399878, or 399952.

In certain embodiments, a target region is nucleotides 601-628 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 601-628 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190,191, 192 or 193. In certain such embodiments, an antisense compoundtargeted to nucleotides 601-628 of SEQ ID NO: 1 is selected from ISISNOs: 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579,410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950,410656, 410535, 410582, 410657, or 406010.

In certain embodiments, a target region is nucleotides 602-628 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 602-628 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191,192 or 193. In certain such embodiments, an antisense compound targetedto nucleotides 602-628 of SEQ ID NO: 1 is selected from ISIS NOs:406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654,410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657,or 406010.

In certain embodiments, a target region is nucleotides 603-630 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 603-630 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192,193 or 194. In certain such embodiments, an antisense compound targetedto nucleotides 603-630 of SEQ ID NO: 1 is selected from ISIS NOs:410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655,399794, 399950, 410656, 410535, 410582, 410657, 406010, or 406011.

In certain embodiments, a target region is nucleotides 611-636 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 611-636 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 13, 194, 195 or 196. Incertain such embodiments, an antisense compound targeted to nucleotides611-636 of SEQ ID NO: 1 is selected from ISIS NOs: 406011, 406012,399795, 399951, or 406013.

In certain embodiments, a target region is nucleotides 620-647 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 620-647 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 14 or 197. In certain suchembodiments, an antisense compound targeted to nucleotides 620-647 ofSEQ ID NO: 1 is selected from ISIS NOs: 395156, 399878, or 410749.

In certain embodiments, a target region is nucleotides 638-665 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 638-665 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain suchembodiments, an antisense compound targeted to nucleotides 638-665 ofSEQ ID NO: 1 is selected from ISIS NOs: 410750, 395157, or 399879.

In certain embodiments, a target region is nucleotides 648-674 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 648-674 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 16, 199, 200, or 201. Incertain such embodiments, an antisense compound targeted to nucleotides648-674 of SEQ ID NO: 1 is selected from ISIS NOs: 406014, 399796,399952, 406015, or 406016.

In certain embodiments, a target region is nucleotides 657-684 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 657-684 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205, or206. In certain such embodiments, an antisense compound targeted tonucleotides 657-684 of SEQ ID NO: 1 is selected from ISIS NOs: 410730,406017, 406018, 406019, or 406020.

In certain embodiments, a target region is nucleotides 705-743 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 705-743 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 17 or 209. In certain suchembodiments, an antisense compound targeted to nucleotides 705-743 ofSEQ ID NO: 1 is selected from ISIS NOs: 395158, 399880, or 410752.

In certain embodiments, a target region is nucleotides 782-810 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 782-810 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213, or214. In certain such embodiments, an antisense compound targeted tonucleotides 782-810 of SEQ ID NO: 1 is selected from ISIS NOs: 406021,406022, 395159, 399881, 406023, 406024, or 410732.

In certain embodiments, a target region is nucleotides 821-859 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 821-859 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217.In certain such embodiments, an antisense compound targeted tonucleotides 821-859 of SEQ ID NO: 1 is selected from ISIS NOs: 410753,406025, 395160, 399882, 406026, 399797, or 399953.

In certain embodiments, a target region is nucleotides 835-859 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 835-859 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20 or 217. In certainsuch embodiments, an antisense compound targeted to nucleotides 835-859of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026,399797, or 399953.

In certain embodiments, a target region is nucleotides 835-917 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 835-917 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, 232 or 233. In certain such embodiments, an antisense compoundtargeted to nucleotides 835-917 of SEQ ID NO: 1 is selected from ISISNOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869,405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876,406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032,395162, 399884, or 410754.

In certain embodiments, a target region is nucleotides 835-942 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 835-942 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24,25, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229,230, 231, 232, or 233. In certain such embodiments, an antisensecompound targeted to nucleotides 835-942 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883,405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875,405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733,406032, 395162, 399884, 410754, 395163, or 399885.

In certain embodiments, a target region is nucleotides 860-887 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 860-887 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220,221, 222 or 223. In certain such embodiments, an antisense compoundtargeted to nucleotides 860-887 of SEQ ID NO: 1 is selected from ISISNOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954,405873, or 405874.

In certain embodiments, a target region is nucleotides 860-899 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 860-899 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 218, 219, 220,221, 222, 223, 224, 225, 226, 227 or 228. In certain such embodiments,an antisense compound targeted to nucleotides 860-899 of SEQ ID NO: 1 isselected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872,399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029,399799, or 399955.

In certain embodiments, a target region is nucleotides 860-909 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 860-909 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219,220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231 or 232. Incertain such embodiments, an antisense compound targeted to nucleotides860-909 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883,405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875,405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733,406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 860-917 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 860-917 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219,220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233.In certain such embodiments, an antisense compound targeted tonucleotides 860-917 of SEQ ID NO: 1 is selected from ISIS NOs: 395161,399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874,405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031,410733, 406032, 395162, 399884, or 410754.

In certain embodiments, a target region is nucleotides 869-895 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 869-895 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 224, 225, 226, or 227. Incertain such embodiments, an antisense compound targeted to nucleotides869-895 of SEQ ID NO: 1 is selected from ISIS NOs: 405875, 405876,406027, or 406028.

In certain embodiments, a target region is nucleotides 878-905 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 878-905 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, or 231.In certain such embodiments, an antisense compound targeted tonucleotides 878-905 of SEQ ID NO: 1 is selected from ISIS NOs: 406029,399799, 399955, 406030, 406031, or 410733.

In certain embodiments, a target region is nucleotides 888-909 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 888-909 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 24 or 232. In certain suchembodiments, an antisense compound targeted to nucleotides 888-909 ofSEQ ID NO: 1 is selected from ISIS NOs: 406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 923-952 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 923-952 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain suchembodiments, an antisense compound targeted to nucleotides 923-952 ofSEQ ID NO: 1 is selected from ISIS NOs: 395163, 399885, or 410755.

In certain embodiments, a target region is nucleotides 960-1034 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 960-1034 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, or 256. In certain such embodiments, an antisensecompound targeted to nucleotides 960-1034 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410756, 405526, 405604, 406033, 395164, 399886, 406034,399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658,410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661,405878, 410587, 410662, 405879, 410588, 410663, 405880, or 410758.

In certain embodiments, a target region is nucleotides 960-1173 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 960-1173 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, or 273. In certain such embodiments,an antisense compound targeted to nucleotides 960-1173 of SEQ ID NO: 1is selected from ISIS NOs: 410756, 405526, 405604, 406033, 395164,399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536,410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877,410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880,or 410763.

In certain embodiments, a target region is nucleotides 960-986 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 960-986 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. Incertain such embodiments, an antisense compound targeted to nucleotides960-986 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526,405604, or 406033.

In certain embodiments, a target region is nucleotides 967-991 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 967-991 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 237 or 238. In certainsuch embodiments, an antisense compound targeted to nucleotides 967-991of SEQ ID NO: 1 is selected from ISIS NOs: 406033 or 406034.

In certain embodiments, a target region is nucleotides 970-1023 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 970-1023 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 238, 239, 240,241, 242, 243, 244, 245, 246, 247, 248, or 249. In certain suchembodiments, an antisense compound targeted to nucleotides 970-1023 ofSEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800,399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537,410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878,410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165,399887, or 410665.

In certain embodiments, a target region is nucleotides 970-1064 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 970-1064 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 238,239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250, 251, 252,253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263. In certainsuch embodiments, an antisense compound targeted to nucleotides 970-1064of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034,399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658,410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661,405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664,395165, 399887, or 399957.

In certain embodiments, a target region is nucleotides 970-1117 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 970-1117 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 31,32, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250,251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264,265, or 266. In certain such embodiments, an antisense compound targetedto nucleotides 970-1117 of SEQ ID NO: 1 is selected from ISIS NOs:395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037,410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660,405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663,405880, 410589, 410664, 395165, 399887, or 399890.

In certain embodiments, a target region is nucleotides 970-996 of SEQ IDNO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 970-996 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, or 238. In certainsuch embodiments, an antisense compound targeted to nucleotides 970-996of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034,399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 977-1004 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 977-1004 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 239 or 240. In certainsuch embodiments, an antisense compound targeted to nucleotides 977-1004of SEQ ID NO: 1 is selected from ISIS NOs: 406035 or 410757.

In certain embodiments, a target region is nucleotides 985-1011 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 985-1011 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. Incertain such embodiments, an antisense compound targeted to nucleotides985-1011 of SEQ ID NO: 1 is selected from ISIS NOs: 410757, 406036, or406037.

In certain embodiments, a target region is nucleotides 989-1016 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 989-1016 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 241, 242, or 243. Incertain such embodiments, an antisense compound targeted to nucleotides989-1016 of SEQ ID NO: 1 is selected from ISIS NOs: 406036, 406037,410536, 410583, or 410658.

In certain embodiments, a target region is nucleotides 992-1019 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 992-1019 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 242, 243, 244, 245, or246. In certain such embodiments, an antisense compound targeted tonucleotides 992-1019 of SEQ ID NO: 1 is selected from ISIS NOs: 406037,410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660,405877, 410586, or 410661.

In certain embodiments, a target region is nucleotides 997-1024 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247,248, 249, or 250. In certain such embodiments, an antisense compoundtargeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISISNOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585,410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588,410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881,410590, or 410666.

In certain embodiments, a target region is nucleotides 997-1024 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247,248, 249, or 250. In certain such embodiments, an antisense compoundtargeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISISNOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585,410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588,410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881,410590, or 410666.

In certain embodiments, a target region is nucleotides 998-1025 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 998-1025 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247,248, 249, 250, or 251. In certain such embodiments, an antisensecompound targeted to nucleotides 998-1025 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877,410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880,410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882,410591, or 410667.

In certain embodiments, a target region is nucleotides 999-1026 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 999-1026 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248,249, 250, 251, or 252. In certain such embodiments, an antisensecompound targeted to nucleotides 999-1026 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 406038, 410585, 410660, 405877, 410586, 410661, 405878,410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165,399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833,410592, or 410668.

In certain embodiments, a target region is nucleotides 1000-1027 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1000-1027 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249,250, 251, 252, or 253. In certain such embodiments, an antisensecompound targeted to nucleotides 1000-1027 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405877, 410586, 410661, 405878, 410587, 410662, 405879,410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881,410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884,410593, or 410669.

In certain embodiments, a target region is nucleotides 1001-1028 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1001-1028 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250,251, 252, 253, or 254. In certain such embodiments, an antisensecompound targeted to nucleotides 1001-1028 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405878, 410587, 410662, 405879, 410588, 410663, 405880,410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882,410591, 410667, 405833, 410592, 410668, 405884, 410593, 410669, 410538,410594, or 410670.

In certain embodiments, a target region is nucleotides 1002-1021 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1002-1021 of SEQ ID NO: 1. In one such embodiment, anantisense compound targeted to nucleotides 1002-1021 of SEQ ID NO: 1 isISIS NO: 405879.

In certain embodiments, a target region is nucleotides 1002-1029 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1002-1029 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251,252, 253, 254, or 255. In certain such embodiments, an antisensecompound targeted to nucleotides 1002-1029 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165,399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833,410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539,410595, or 410671.

In certain embodiments, a target region is nucleotides 1003-1029 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1003-1029 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252,253, 254, or 255. In certain such embodiments, an antisense compoundtargeted to nucleotides 1003-1029 of SEQ ID NO: 1 is selected from ISISNOs: 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881,410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884,410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1004-1029 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1004-1029 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253,254, or 255. In certain such embodiments, an antisense compound targetedto nucleotides 1004-1029 of SEQ ID NO: 1 is selected from ISIS NOs:395165, 399887, 409126, 410665, 405881, 410590, 410666, 405882, 410591,410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594,410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1005-1029 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1005-1029 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254,or 255. In certain such embodiments, an antisense compound targeted tonucleotides 1005-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 410665,405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668,405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or410671.

In certain embodiments, a target region is nucleotides 1006-1029 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1006-1029 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or255. In certain such embodiments, an antisense compound targeted tonucleotides 1006-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405882,410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538,410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1007-1034 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1007-1034 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255, or256. In certain such embodiments, an antisense compound targeted tonucleotides 1007-1034 of SEQ ID NO: 1 is selected from ISIS NOs: 405883,410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539,410595, 410671, or 410758.

In certain embodiments, a target region is nucleotides 1036-1061 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1036-1061 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 29, 257, 258, or 259. Incertain such embodiments, an antisense compound targeted to nucleotides1036-1061 of SEQ ID NO: 1 is selected from ISIS NOs: 406039, 406040,395166, 399888, or 406041.

In certain embodiments, a target region is nucleotides 1045-1072 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1045-1072 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 30, 260, 261, or 262. Incertain such embodiments, an antisense compound targeted to nucleotides1045-1072 of SEQ ID NO: 1 is selected from ISIS NOs: 399801, 399957,406042, 406043, or 406044.

In certain embodiments, a target region is nucleotides 1076-1096 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1076-1096 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 31 or 264. In certain suchembodiments, an antisense compound targeted to nucleotides 1076-1096 ofSEQ ID NO: 1 is selected from ISIS NOs: 408642, 395167, or 399889.

In certain embodiments, a target region is nucleotides 1088-1115 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1088-1115 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 265 or 266. In certainsuch embodiments, an antisense compound targeted to nucleotides1088-1115 of SEQ ID NO: 1 is selected from ISIS NOs: 410760 or 406045.

In certain embodiments, a target region is nucleotides 1098-1123 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1098-1123 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 32, 267, 268, or 269. Incertain such embodiments, an antisense compound targeted to nucleotides1098-1123 of SEQ ID NO: 1 is selected from ISIS NOs: 395168, 399890,405909, 405910, or 405911.

In certain embodiments, a target region is nucleotides 1200-1251 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1200-1251 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, or 285. In certain such embodiments,an antisense compound targeted to nucleotides 1200-1251 of SEQ ID NO: 1is selected from ISIS NOs: 410764, 395169, 399891, 405913, 405914,405915, 405916, 410734, 405917, 405918, 405919, 405920, 405921, or405922.

In certain embodiments, a target region is nucleotides 1210-1237 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1210-1237 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278.In certain such embodiments, an antisense compound targeted tonucleotides 1210-1237 of SEQ ID NO: 1 is selected from ISIS NOs: 395169,399891, 405913, 405914, 405915, or 405916.

In certain embodiments, a target region is nucleotides 1219-1245 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1219-1245 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. Incertain such embodiments, an antisense compound targeted to nucleotides1219-1245 of SEQ ID NO: 1 is selected from ISIS NOs: 410734, 405917,405918, or 405919.

In certain embodiments, a target region is nucleotides 1228-1251 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1228-1251 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. Incertain such embodiments, an antisense compound targeted to nucleotides1228-1251 of SEQ ID NO: 1 is selected from ISIS NOs: 405920, 405921, or405922.

In certain embodiments, a target region is nucleotides 1273-1444 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1273-1444 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39,40, 41, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298,299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312,313, 314, 315, 316, 316, and 318. In certain such embodiments, anantisense compound targeted to nucleotides 1273-1444 of SEQ ID NO: 1 isselected from ISIS NOs: 410765, 410766, 405923, 395170, 399892, 410767,405924, 410735, 405925, 405926, 395171, 399893, 405927, 395172, 399894,405928, 399802, 399958, 405929, 395173, 399895, 405930, 405931, 405932,405933, 405934, 399803, 399959, 405935, 410736, 405936, 395174, or410769.

In certain embodiments, a target region is nucleotides 1295-1316 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1295-1316 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 149 or 288. In certainsuch embodiments, an antisense compound targeted to nucleotides1295-1316 of SEQ ID NO: 1 is selected from ISIS NOs: 405923, 395170, or399892.

In certain embodiments, a target region is nucleotides 1318-1345 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1318-1345 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293.In certain such embodiments, an antisense compound targeted tonucleotides 1318-1345 of SEQ ID NO: 1 is selected from ISIS NOs: 405924,410735, 405925, 405926, 395171, or 399893.

In certain embodiments, a target region is nucleotides 1328-1354 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1328-1354 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 35, 128, 294 or 295. Incertain such embodiments, an antisense compound targeted to nucleotides1328-1354 of SEQ ID NO: 1 is selected from ISIS NOs: 405927, 395172,399894, 405928, 399802, or 399958.

In certain embodiments, a target region is nucleotides 1337-1361 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1337-1361 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 36, 296, or 297. Incertain such embodiments, an antisense compound targeted to nucleotides1337-1361 of SEQ ID NO: 1 is selected from ISIS NOs: 405929, 395173,399895, or 405930.

In certain embodiments, a target region is nucleotides 1344-1371 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1344-1371 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 37, 298, 299, 300, or 301.In certain such embodiments, an antisense compound targeted tonucleotides 1344-1371 of SEQ ID NO: 1 is selected from ISIS NOs: 405931,405932, 405933, 405934, 399803, or 399959.

In certain embodiments, a target region is nucleotides 1354-1377 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1354-1377 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 302, 303, or 304. Incertain such embodiments, an antisense compound targeted to nucleotides1354-1377 of SEQ ID NO: 1 is selected from ISIS NOs: 405935, 410736, or405936.

In certain embodiments, a target region is nucleotides 1380-1406 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1380-1406 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 305 or 306. In certainsuch embodiments, an antisense compound targeted to nucleotides1380-1406 of SEQ ID NO: 1 is selected from ISIS NOs: 410768 or 405937.

In certain embodiments, a target region is nucleotides 1389-1416 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1389-1416 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310.In certain such embodiments, an antisense compound targeted tonucleotides 1389-1416 of SEQ ID NO: 1 is selected from ISIS NOs: 395175,399897, 405938, 405939, 405940, or 405941.

In certain embodiments, a target region is nucleotides 1400-1426 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1400-1426 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313, or 314.In certain such embodiments, an antisense compound targeted tonucleotides 1400-1426 of SEQ ID NO: 1 is selected from ISIS NOs: 399804,399960, 405942, 405943, 410737, or 405944.

In certain embodiments, a target region is nucleotides 1409-1434 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1409-1434 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 41, 315, 316, or 317. Incertain such embodiments, an antisense compound targeted to nucleotides1409-1434 of SEQ ID NO: 1 is selected from ISIS NOs: 405945, 399805,399961, 405946, or 405947.

In certain embodiments, a target region is nucleotides 1465-1491 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1465-1491 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. Incertain such embodiments, an antisense compound targeted to nucleotides1465-1491 of SEQ ID NO: 1 is selected from ISIS NOs: 395176, 399898,405948, 399806, 399962, or 405949.

In certain embodiments, a target region is nucleotides 1465-1602 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1465-1602 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327,328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341,342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356,357, 358, or 359. In certain such embodiments, an antisense compoundtargeted to nucleotides 1465-1602 of SEQ ID NO: 1 is selected from ISISNOs: 395176, 399898, 405948, 399806, 399962, 405949, 405950, 405951,399807, 399963, 405952, 410738, 405953, 405954, 410770, 405955, 405956,405957, 405958, 405959, 405960, 405961, 399808, 399964, 405962, 410739,405963, 395177, 399899, 405964, 399809, 399965, or 399969.

In certain embodiments, a target region is nucleotides 1474-1499 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1474-1499 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 321, 322, or 323. Incertain such embodiments, an antisense compound targeted to nucleotides1474-1499 of SEQ ID NO: 1 is selected from ISIS NOs: 405950, 405951,399807, 399963, or 405952.

In certain embodiments, a target region is nucleotides 1482-1519 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1482-1519 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 324, 325, 326, or 327. Incertain such embodiments, an antisense compound targeted to nucleotides1482-1519 of SEQ ID NO: 1 is selected from ISIS NOs: 410738, 405953,405954, or 410770.

In certain embodiments, a target region is nucleotides 1513-1540 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1513-1540 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331, or332. In certain such embodiments, an antisense compound targeted tonucleotides 1513-1540 of SEQ ID NO: 1 is selected from ISIS NOs: 405955,405956, 405957, 405958, or 405959.

In certain embodiments, a target region is nucleotides 1523-1549 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1523-1549 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335, or 336.In certain such embodiments, an antisense compound targeted tonucleotides 1523-1549 of SEQ ID NO: 1 is selected from ISIS NOs: 405960,405961, 399808, 399964, 405962, or 410739.

In certain embodiments, a target region is nucleotides 1526-1602 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1526-1602 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49,50, 51, 87, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347,348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. Incertain such embodiments, an antisense compound targeted to nucleotides1526-1602 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964,405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965,405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970,410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673,399812, 399968, 410674, or 399969.

In certain embodiments, a target region is nucleotides 1526-1624 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1526-1624 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49,50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346,347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. Incertain such embodiments, an antisense compound targeted to nucleotides1526-1624 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964,405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965,405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970,410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673,399812, 399968, 410674, or 399902.

In certain embodiments, a target region is nucleotides 1532-1558 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1532-1558 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 45, 46, 337, or 338. Incertain such embodiments, an antisense compound targeted to nucleotides1532-1558 of SEQ ID NO: 1 is selected from ISIS NOs: 405963, 395177,399899, 405964, 399809, or 399965.

In certain embodiments, a target region is nucleotides 1541-1568 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1541-1568 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 47, 340, 341, or 342. Incertain such embodiments, an antisense compound targeted to nucleotides1541-1568 of SEQ ID NO: 1 is selected from ISIS NOs: 405965, 405966,399810, 399966, 405967, or 405968.

In certain embodiments, a target region is nucleotides 1552-1579 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1552-1579 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 48, 343, 344, 345, or 346.In certain such embodiments, an antisense compound targeted tonucleotides 1552-1579 of SEQ ID NO: 1 is selected from ISIS NOs: 399811,399967, 405969, 405970, 410740, or 405885.

In certain embodiments, a target region is nucleotides 1560-1587 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1560-1587 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349,350, 351, 352, or 353. In certain such embodiments, an antisensecompound targeted to nucleotides 1560-1587 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405885, 405886, 405887, 410596, 410672, 405888, 410597,410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599,410676, 405891, 410600, 410677, 405892, 410601, or 410678.

In certain embodiments, a target region is nucleotides 1561-1589 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1561-1589 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 346, 347, 348,349, 350, 351, 352, 353, or 354. In certain such embodiments, anantisense compound targeted to nucleotides 1561-1589 of SEQ ID NO: 1 isselected from ISIS NOs: 405886, 405887, 410596, 410672, 405888, 410597,410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599,410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900,405557, 410679, 408653, 410602, or 410680.

In certain embodiments, a target region is nucleotides 1564-1591 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1564-1591 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352,353, 354, 355, or 356. In certain such embodiments, an antisensecompound targeted to nucleotides 1564-1591 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890,410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178,399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681,410540, 410604, or 410682.

In certain embodiments, a target region is nucleotides 1565-1592 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1565-1592 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353,354, 355, 356, or 357. In certain such embodiments, an antisensecompound targeted to nucleotides 1565-1592 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891,410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679,408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682,or 405972.

In certain embodiments, a target region is nucleotides 1566-1592 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1566-1592 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354,355, 356, or 357. In certain such embodiments, an antisense compoundtargeted to nucleotides 1566-1592 of SEQ ID NO: 1 is selected from ISISNOs: 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601,410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971,410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1567-1592 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1567-1592 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 352, 353, 354, 355,356, or 357. In certain such embodiments, an antisense compound targetedto nucleotides 1567-1592 of SEQ ID NO: 1 is selected from ISIS NOs:405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557,410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604,410682, or 405972.

In certain embodiments, a target region is nucleotides 1570-1597 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1570-1597 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 354, 355, 356, 357 or 358.In certain such embodiments, an antisense compound targeted tonucleotides 1570-1597 of SEQ ID NO: 1 is selected from ISIS NOs: 408653,410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, 405972or 405973.

In certain embodiments, a target region is nucleotides 1571-1599 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1571-1599 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 87, 356, 357, 358, or 359.In certain such embodiments, an antisense compound targeted tonucleotides 1571-1599 of SEQ ID NO: 1 is selected from ISIS NOs: 405971,410603, 410681, 410540, 410604, 410682, 405972, 395179, 399901, 405973,or 405974.

In certain embodiments, a target region is nucleotides 1605-1706 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1605-1706 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 119, 360, 361,362, 363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain suchembodiments, an antisense compound targeted to nucleotides 1605-1706 ofSEQ ID NO: 1 is selected from ISIS NOs: 395180, 399902, 410771, 395181,399903, 405975, 399814, 399970, 405976, 405977, 410772, 405978, 395182,399904, 405979, 405980, 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 1628-1706 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1628-1706 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 360, 361, 362,363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain suchembodiments, an antisense compound targeted to nucleotides 1628-1706 ofSEQ ID NO: 1 is selected from ISIS NOs: 410771, 395181, 399903, 405975,399814, 399970, 405976, 405977, 410772, 405978, 395182, 399904, 405979,405980, 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 1640-1666 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1640-1666 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. Incertain such embodiments, an antisense compound targeted to nucleotides1640-1666 of SEQ ID NO: 1 is selected from ISIS NOs: 395181, 399903,405975, 399814, 399970, or 405976.

In certain embodiments, a target region is nucleotides 1672-1698 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1672-1698 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. Incertain such embodiments, an antisense compound targeted to nucleotides1672-1698 of SEQ ID NO: 1 is selected from ISIS NOs: 405978, 395182,399904, 405979, or 405980.

In certain embodiments, a target region is nucleotides 1681-1706 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1681-1706 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 368, 369, 370, or 371. Incertain such embodiments, an antisense compound targeted to nucleotides1681-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 405981, 410741,405982, or 405983.

In certain embodiments, a target region is nucleotides 1735-1761 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1735-1761 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 372, 373, or 374. Incertain such embodiments, an antisense compound targeted to nucleotides1735-1761 of SEQ ID NO: 1 is selected from ISIS NOs: 405984, 405564,405641, 405985, 399815, 399971, or 405986.

In certain embodiments, a target region is nucleotides 1735-1765 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1735-1765 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or375. In certain such embodiments, an antisense compound targeted tonucleotides 1735-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 405984,405564, 405641, 405985, 399815, 399971, 405986, 405987, 399816 or399972.

In certain embodiments, a target region is nucleotides 1740-1765 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1740-1765 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 56, 374, or 375. Incertain such embodiments, an antisense compound targeted to nucleotides1740-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 399815, 399971,405986, 405987, 399816, or 399972.

In certain embodiments, a target region is nucleotides 1849-1876 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1849-1876 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380,381, 382, 383, or 384. In certain such embodiments, an antisensecompound targeted to nucleotides 1849-1876 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684, 410543,410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988,410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547,410613, or 410691.

In certain embodiments, a target region is nucleotides 1849-1879 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1849-1879 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379,380, 381, 382, 383, 384, 385, or 386. In certain such embodiments, anantisense compound targeted to nucleotides 1849-1879 of SEQ ID NO: 1 isselected from ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684,410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687,405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690,410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, or405990.

In certain embodiments, a target region is nucleotides 1850-1877 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1850-1877 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380,381, 382, 383 or 384. In certain such embodiments, an antisense compoundtargeted to nucleotides 1850-1877 of SEQ ID NO: 1 is selected from ISISNOs: 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608,410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611,410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906,or 410692.

In certain embodiments, a target region is nucleotides 1851-1877 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1851-1877 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381,382, 383, or 384. In certain such embodiments, an antisense compoundtargeted to nucleotides 1851-1877 of SEQ ID NO: 1 is selected from ISISNOs: 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609,410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612,410690, 410547, 410613, 395184, 410691, 399906, or 410692.

In certain embodiments, a target region is nucleotides 1852-1878 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1852-1878 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382,383, 384, or 385. In certain such embodiments, an antisense compoundtargeted to nucleotides 1852-1878 of SEQ ID NO: 1 is selected from ISISNOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610,410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613,395184, 410691, 399906, 410692, 410548, 410614, or 410693.

In certain embodiments, a target region is nucleotides 1852-1879 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1852-1879 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382,383, 384, 385, or 386. In certain such embodiments, an antisensecompound targeted to nucleotides 1852-1879 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988,410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547,410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or405990.

In certain embodiments, a target region is nucleotides 1853-1879 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1853-1879 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383,384, 385 or 386. In certain such embodiments, an antisense compoundtargeted to nucleotides 1853-1879 of SEQ ID NO: 1 is selected from ISISNOs: 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611,410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906,410692, 410548, 410614, 410693, or 405990.

In certain embodiments, a target region is nucleotides 1854-1879 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1854-1879 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384,385, or 386. In certain such embodiments, an antisense compound targetedto nucleotides 1854-1879 of SEQ ID NO: 1 is selected from ISIS NOs:405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690,410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693,or 405990.

In certain embodiments, a target region is nucleotides 1905-1955 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1905-1955 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 387, 388, 389, 390,391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404,405, or 406. In certain such embodiments, an antisense compound targetedto nucleotides 1905-1955 of SEQ ID NO: 1 is selected from ISIS NOs:410773, 410549, 410615, 410694, 410550, 410616, 410695, 410551, 410617,410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907,410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622,410702, 405992, 410623, 410703, 410556, or 410774.

In certain embodiments, a target region is nucleotides 1915-1942 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1915-1942 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391,392, 393, 394, or 395. In certain such embodiments, an antisensecompound targeted to nucleotides 1915-1942 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410549, 410615, 410694, 410550, 410616, 410695, 410551,410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185,399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555,410622, or 410702.

In certain embodiments, a target region is nucleotides 1916-1943 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1916-1943 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392,393, 394, 395, or 396. In certain such embodiments, an antisensecompound targeted to nucleotides 1916-1943 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410550, 410616, 410695, 410551, 410617, 410696, 410552,410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554,410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992,410623, or 410703.

In certain embodiments, a target region is nucleotides 1917-1944 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1917-1944 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393,394, 395, 396, or 397. In certain such embodiments, an antisensecompound targeted to nucleotides 1917-1944 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410551, 410617, 410696, 410552, 410618, 410697, 410553,410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991,410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556,410624, or 410704.

In certain embodiments, a target region is nucleotides 1918-1945 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1918-1945 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394,395, 396, 397, or 398. In certain such embodiments, an antisensecompound targeted to nucleotides 1918-1945 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410552, 410618, 410697, 410553, 410619, 410698, 395185,399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555,410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993,410625, or 410705.

In certain embodiments, a target region is nucleotides 1919-1946 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1919-1946 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395,396, 397, or 399. In certain such embodiments, an antisense compoundtargeted to nucleotides 1919-1946 of SEQ ID NO: 1 is selected from ISISNOs: 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620,410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623,410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626,or 410706.

In certain embodiments, a target region is nucleotides 1920-1939 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1920-1939 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59. In certain suchembodiments, an antisense compound targeted to nucleotides 1920-1939 ofSEQ ID NO: 1 is selected from ISIS NOs: 395185, 399907, or 410699.

In certain embodiments, a target region is nucleotides 1920-1947 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1920-1947 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396,397, 398, 399, or 400. In certain such embodiments, an antisensecompound targeted to nucleotides 1920-1947 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 395185, 399907, 410699, 410554, 410620, 410700, 405991,410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556,410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944,410627, or 410707.

In certain embodiments, a target region is nucleotides 1921-1948 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1921-1948 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397,398, 399, 400, or 401. In certain such embodiments, an antisensecompound targeted to nucleotides 1921-1948 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410554, 410620, 410700, 405991, 410621, 410701, 410555,410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993,410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558,410628, or 410708.

In certain embodiments, a target region is nucleotides 1922-1949 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1922-1949 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398,399, 400, 401, or 402. In certain such embodiments, an antisensecompound targeted to nucleotides 1922-1949 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405991, 410621, 410701, 410555, 410622, 410702, 405992,410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557,410626, 410706, 405944, 410627, 410707, 410558, 410628, 410708, 405995,410629, or 410709.

In certain embodiments, a target region is nucleotides 1923-1950 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1923-1950 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399,400, 401, 402, or 403. In certain such embodiments, an antisensecompound targeted to nucleotides 1923-1950 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410555, 410622, 410702, 405992, 410623, 410703, 410556,410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944,410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559,410630, or 410710.

In certain embodiments, a target region is nucleotides 1924-1951 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1924-1951 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400,401, 402, 403, or 404. In certain such embodiments, an antisensecompound targeted to nucleotides 1924-1951 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 405992, 410623, 410703, 410556, 410624, 410704, 405993,410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558,410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560,410631, or 410711.

In certain embodiments, a target region is nucleotides 1925-1952 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1925-1952 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401,402, 403, 404, or 405. In certain such embodiments, an antisensecompound targeted to nucleotides 1925-1952 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410556, 410624, 410704, 405993, 410625, 410705, 410557,410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995,410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561,410632, or 410712.

In certain embodiments, a target region is nucleotides 1926-1952 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1926-1952 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402,403, 404, or 405. In certain such embodiments, an antisense compoundtargeted to nucleotides 1926-1952 of SEQ ID NO: 1 is selected from ISISNOs: 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627,410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630,410710, 410560, 410631, 410711, 410561, 410632, or 410712.

In certain embodiments, a target region is nucleotides 1927-1952 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1927-1952 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403,404, or 405. In certain such embodiments, an antisense compound targetedto nucleotides 1927-1952 of SEQ ID NO: 1 is selected from ISIS NOs:410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708,405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711,410561, 410632, or 410712.

In certain embodiments, a target region is nucleotides 1928-1955 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1928-1955 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404,405, or 406. In certain such embodiments, an antisense compound targetedto nucleotides 1928-1955 of SEQ ID NO: 1 is selected from ISIS NOs:405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709,410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, 410712,or 410774.

In certain embodiments, a target region is nucleotides 1962-2059 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 1962-2059 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 407, 408, 409, or 410. Incertain such embodiments, an antisense compound targeted to nucleotides1962-2059 of SEQ ID NO: 1 is selected from ISIS NOs: 410775, 410776,410777, or 410778.

In certain embodiments, a target region is nucleotides 2040-2126 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2040-2126 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412,413, 414, or 415. In certain such embodiments, an antisense compoundtargeted to nucleotides 2040-2126 of SEQ ID NO: 1 is selected from ISISNOs: 410778, 395186, 399908, 410713, 410562, 410633, 410714, 405996,410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817,399973, 410718, or 405997.

In certain embodiments, a target region is nucleotides 2100-2126 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2100-2126 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413,414, or 415. In certain such embodiments, an antisense compound targetedto nucleotides 2100-2126 of SEQ ID NO: 1 is selected from ISIS NOs:395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715,410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718or 405997.

In certain embodiments, a target region is nucleotides 2100-2139 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2100-2139 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413,414, 415, or 416. In certain such embodiments, an antisense compoundtargeted to nucleotides 2100-2139 of SEQ ID NO: 1 is selected from ISISNOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634,410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973,410718, 405997 or 410779.

In certain embodiments, a target region is nucleotides 2100-2206 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2100-2206 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413,414, 415, 416, 417, or 418. In certain such embodiments, an antisensecompound targeted to nucleotides 2100-2206 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996,410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817,399973, 410718, 405997, 405997, 410780, or 410781.

In certain embodiments, a target region is nucleotides 2101-2126 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2101-2126 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 61, 411, 412, 413, 414, or415. In certain such embodiments, an antisense compound targeted tonucleotides 2101-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 410562,410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564,410636, 410717, 399817, 399973, 410718, or 405997.

In certain embodiments, a target region is nucleotides 2305-2332 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2305-2332 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425, or 426. In certain such embodiments, an antisensecompound targeted to nucleotides 2305-2332 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410565, 410637, 410719, 410566, 410638, 410720, 410567,410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187,399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998,410644, or 410727.

In certain embodiments, a target region is nucleotides 2305-2354 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2305-2354 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments,an antisense compound targeted to nucleotides 2305-2354 of SEQ ID NO: 1is selected from ISIS NOs: 410565, 410637, 410719, 410566, 410638,410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641,410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643,410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646,or 410783.

In certain embodiments, a target region is nucleotides 2306-2333 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2306-2333 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423,424, 425, 426, or 427. In certain such embodiments, an antisensecompound targeted to nucleotides 2306-2333 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568,410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570,410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572,410645, or 410728.

In certain embodiments, a target region is nucleotides 2307-2334 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2307-2334 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424,425, 426, 427, or 428. In certain such embodiments, an antisensecompound targeted to nucleotides 2307-2334 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 410567, 410639, 410721, 410568, 410640, 410722, 410569,410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571,410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573,410646, or 410729.

In certain embodiments, a target region is nucleotides 2308-2334 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2308-2334 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425,426, 427, or 428. In certain such embodiments, an antisense compoundtargeted to nucleotides 2308-2334 of SEQ ID NO: 1 is selected from ISISNOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909,410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644,410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2309-2334 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2309-2334 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426,427, or 428. In certain such embodiments, an antisense compound targetedto nucleotides 2309-2334 of SEQ ID NO: 1 is selected from ISIS NOs:410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725,410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728,410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2310-2334 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2310-2334 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427, or428. In certain such embodiments, an antisense compound targeted tonucleotides 2310-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 395187,399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998,410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2410-2434 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2410-2434 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain suchembodiments, an antisense compound targeted to nucleotides 2410-2434 ofSEQ ID NO: 1 is selected from ISIS NOs: 395188, 399910, 399818, or399974.

In certain embodiments, a target region is nucleotides 2504-2528 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2504-2528 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain suchembodiments, an antisense compound targeted to nucleotides 2504-2528 ofSEQ ID NO: 1 is selected from ISIS NOs: 395189, 399911, 399819, or399975.

In certain embodiments, a target region is nucleotides 2509-2528 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2509-2528 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 65. In certain suchembodiments, an antisense compound targeted to nucleotides 2509-2528 ofSEQ ID NO: 1 is selected from ISIS NOs: 399819 or 399975.

In certain embodiments, a target region is nucleotides 2582-2625 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2582-2625 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certainsuch embodiments, an antisense compound targeted to nucleotides2582-2625 of SEQ ID NO: 1 is selected from ISIS NOs: 399820, 399976,395190, 399912, 395191, or 399913.

In certain embodiments, a target region is nucleotides 2606-2668 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2606-2668 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 67 or 153. In certain suchembodiments, an antisense compound targeted to nucleotides 2606-2668 ofSEQ ID NO: 1 is selected from ISIS NOs: 395191, 399913, 395192, or399914.

In certain embodiments, a target region is nucleotides 2828-2855 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2828-2855 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434,435, 436, 437 or 438. In certain such embodiments, an antisense compoundtargeted to nucleotides 2828-2855 of SEQ ID NO: 1 is selected from ISISNOs: 405893, 405894, 405895, 405896, 395194, 399916, 405897, 405898,405899, or 405900.

In certain embodiments, a target region is nucleotides 2832-2851 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2832-2851 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 69. In certain suchembodiments, an antisense compound targeted to nucleotides 2832-2851 ofSEQ ID NO: 1 is selected from ISIS NOs: 395194, or 399916.

In certain embodiments, a target region is nucleotides 2900-2927 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2900-2927 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441,442, 443 or 444. In certain such embodiments, an antisense compoundtargeted to nucleotides 2900-2927 of SEQ ID NO: 1 is selected from ISISNOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821, 399977,405905, or 405906.

In certain embodiments, a target region is nucleotides 2900-2929 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2900-2929 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441,442, 443, 444, 446 or 446. In certain such embodiments, an antisensecompound targeted to nucleotides 2900-2929 of SEQ ID NO: 1 is selectedfrom ISIS NOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821,399977, 405905, 405906, 405907, or 405908.

In certain embodiments, a target region is nucleotides 2902-2927 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2902-2927 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 71, 439, 440, 441, 442,443 or 444. In certain such embodiments, an antisense compound targetedto nucleotides 2902-2927 of SEQ ID NO: 1 is selected from ISIS NOs:405901, 405902, 405903, 405904, 399821, 399977, 405905, or 405906.

In certain embodiments, a target region is nucleotides 2983-3007 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2983-3007 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 72 or 73. In certain suchembodiments, an antisense compound targeted to nucleotides 2983-3007 ofSEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918, 399822, or399978.

In certain embodiments, a target region is nucleotides 2983-3013 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 2983-3013 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certainsuch embodiments, an antisense compound targeted to nucleotides2983-3013 of SEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918,399822, 399978, 399823, or 399979.

In certain embodiments, a target region is nucleotides 3227-3252 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 3227-3252 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 74 or 112. In certain suchembodiments, an antisense compound targeted to nucleotides 3227-3252 ofSEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919, 399824, or399980.

In certain embodiments, a target region is nucleotides 3227-3456 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 3227-3456 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certainsuch embodiments, an antisense compound targeted to nucleotides3227-3456 of SEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919,399824, 399980, 395198, or 399920.

In certain embodiments, a target region is nucleotides 3472-3496 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 3472-3496 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain suchembodiments, an antisense compound targeted to nucleotides 3472-3496 ofSEQ ID NO: 1 is selected from ISIS NOs: 395199, 399921, 399825 or399981.

In certain embodiments, a target region is nucleotides 3543-3569 of SEQID NO: 1. In certain embodiments, an antisense compound is targeted tonucleotides 3543-3569 of SEQ ID NO: 1. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain suchembodiments, an antisense compound targeted to nucleotides 3543-3569 ofSEQ ID NO: 1 is selected from ISIS NOs: 395200, 399922, 399826 or399982.

In certain embodiments, antisense compound target a PCSK9 nucleic acidhaving the sequence of nucleotides 25475000 to 25504000 of GENBANK®Accession No. NT_032977.8, first deposited with GENBANK® on Feb. 26,2006, and incorporated herein as SEQ ID NO: 2. In certain suchembodiments, an antisense oligonucleotide is targeted to SEQ ID NO: 2.In certain such embodiments, an antisense oligonucleotide that istargeted to SEQ ID NO: 2 is at least 90% complementary to SEQ ID NO: 2.In certain such embodiments, an antisense oligonucleotide that istargeted to SEQ ID NO: 2 is at least 95% complementary to SEQ ID NO: 2.In certain such embodiments, an antisense oligonucleotide that istargeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. Incertain such embodiments, an antisense oligonucleotide comprises anucleotide sequence selected from a nucleotide sequence set forth inTable 7.

TABLE 7 Antisense sequences targeted to NT_032977.8 (SEQ ID NO: 2)5′Start 3′Start Site to Site to SEQ ID SEQ ID SEQ ID NO NO: 2 NO: 2Sequence (5′to 3′) 4 2274 2293 GCGCGGAATCCTGGCTGGGA 5 2381 2400GAGGAGACCTAGAGGCCGTG 159 2433 2452 GCCTGGAGCTGACGGTGCCC 160 2437 2456GACCGCCTGGAGCTGACGGT 6 2439 2458 AGGACCGCCTGGAGCTGACG 162 2545 2564AAGGCTAGCACCAGCTCCTC 163 2546 2565 CAAGGCTAGCACCAGCTCCT 164 2547 2566GCAAGGCTAGCACCAGCTCC 165 2548 2567 CGCAAGGCTAGCACCAGCTC 7 2549 2568ACGCAAGGCTAGCACCAGCT 166 2550 2569 AACGCAAGGCTAGCACCAGC 167 2551 2570GAACGCAAGGCTAGCACCAG 168 2552 2571 GGAACGCAAGGCTAGCACCA 169 2553 2572CGGAACGCAAGGCTAGCACC 8 2556 2575 CCTCGGAACGCAAGGCTAGC 170 2560 2579TCCTCCTCGGAACGCAAGGC 171 2585 2604 GTGCTCGGGTGCTTCGGCCA 172 2605 2624TGGAAGGTGGCTGTGGTTCC 9 2619 2638 CCTTGGCGCAGCGGTGGAAG 107 3056 3075CCCACTATAATGGCAAGCCC 80 4306 4325 AACCCAGTTCTAATGCACCT 106 5140 5159CCAGTCAGAGTAGAACAGAG 102 5590 5609 ATGTGCAGAGATCAATCACA 121 5599 5618GGAGCCTACATGTGCAGAGA 94 5667 5686 AGCATGGCACCAGCATCTGC 176 6444 6463CGTAGGTGCCAGGCAACCTC 177 6482 6501 TGACTGCGAGAGGTGGGTCT 178 6492 6511CAGTGCGCTCTGACTGCGAG 179 6494 6513 GGCAGTGCGCTCTGACTGCG 180 6496 6515CGGGCAGTGCGCTCTGACTG 10 6498 6517 GGCGGGCAGTGCGCTCTGAC 181 6499 6518CGGCGGGCAGTGCGCTCTGA 182 6528 6547 ATCCCCGGCGGGCAGCCTGG 183 6532 6551AGGTATCCCCGGCGGGCAGC 184 6534 6553 TGAGGTATCCCCGGCGGGCA 185 6535 6554GTGAGGTATCCCCGGCGGGC 186 6536 6555 GGTGAGGTATCCCCGGCGGG 11 6537 6556TGGTGAGGTATCCCCGGCGG 187 6538 6557 TTGGTGAGGTATCCCCGGCG 188 6539 6558CTTGGTGAGGTATCCCCGGC 189 6540 6559 TCTTGGTGAGGTATCCCCGG 190 6541 6560ATCTTGGTGAGGTATCCCCG 191 6542 6561 GATCTTGGTGAGGTATCCCC 12 6543 6562GGATCTTGGTGAGGTATCCC 192 6544 6563 AGGATCTTGGTGAGGTATCC 193 6546 6565GCAGGATCTTGGTGAGGTAT 194 6548 6567 ATGCAGGATCTTGGTGAGGT 195 6550 6569ACATGCAGGATCTTGGTGAG 13 6552 6571 AGACATGCAGGATCTTGGTG 196 6554 6573GAAGACATGCAGGATCTTGG 14 6557 6576 ATGGAAGACATGCAGGATCT 197 6565 6584AGAAGGCCATGGAAGACATG 198 6575 6594 GAAGCCAGGAAGAAGGCCAT 15 6583 6602TTCACCAGGAAGCCAGGAAG 199 6585 6604 TCTTCACCAGGAAGCCAGGA 16 6588 6607TCATCTTCACCAGGAAGCCA 200 6590 6609 ACTCATCTTCACCAGGAAGC 201 6592 6611CCACTCATCTTCACCAGGAA 202 6594 6613 CGCCACTCATCTTCACCAGG 203 6596 6615GTCGCCACTCATCTTCACCA 204 6598 6617 AGGTCGCCACTCATCTTCAC 205 6600 6619GCAGGTCGCCACTCATCTTC 206 6602 6621 CAGCAGGTCGCCACTCATCT 207 6604 6623TCCAGCAGGTCGCCACTCAT 108 6652 6671 CCCAGCCCTATCAGGAAGTG 144 7099 7118TGACATCCAGGAGGGAGGAG 91 7556 7575 AGACTGATGGAAGGCATTGA 131 7565 7584GTGTTGAGCAGACTGATGGA 145 8836 8855 TGACATCTTGTCTGGGAGCC 90 8948 8967AGACTAGGAGCCTGAGTTTT 125 9099 9118 GGCCTGCAGAAGCCAGAGAG 17 9130 9149CCTCGATGTAGTCGACATGG 209 9149 9168 GCAAAGACAGAGGAGTCCTC 210 9207 9226TTCATCCGCCCGGTACCGTG 211 9209 9228 TATTCATCCGCCCGGTACCG 18 9210 9229GTATTCATCCGCCCGGTACC 212 9212 9231 TGGTATTCATCCGCCCGGTA 213 9214 9233GCTGGTATTCATCCGCCCGG 214 9216 9235 GGGCTGGTATTCATCCGCCC 148 10252 10271TGGCAGCAACTCAGACATAT 127 10633 10652 GGTGGTAATTTGTCACAGCA 84 11308 11327AAGGTCACACAGTTAAGAGT 79 11472 11491 AAATGCAGGGCTAAAATCAC 88 12715 12734ACTGGATACATTGGCAGACA 111 12928 12947 CTAGAGGAACCACTAGATAT 85 13681 13700ACAAATTCCCAGACTCAGCA 100 13746 13765 ATCTCAGGACAGGTGAGCAA 116 1376013779 GAGTAGAGATTCTCATCTCA 129 13816 13835 GTGCCATCTGAACAGCACCT 11713828 13847 GAGTCTTCTGAAGTGCCATC 81 13903 13922 AAGCAGGGCCTCAGGTGGAA 11013926 13945 CCTGGAACCCCTGCAGCCAG 152 13977 13996 TTCAGGCAGGTTGCTGCTAG 8313986 14005 AAGGAAGACTTCAGGCAGGT 140 13998 14017 TCAGCCAGGCCAAAGGAAGA137 14112 14131 TAGGGAGAGCTCACAGATGC 136 14122 14141TAGGAGAAAGTAGGGAGAGC 132 14179 14198 TAAAAGCTGCAAGAGACTCA 139 1426714286 TCAGAGAAAACAGTCACCGA 92 14397 14416 AGAGACAGGAAGCTGCAGCT 142 1440414423 TCATTTTAGAGACAGGAAGC 113 14441 14460 GAATAACAGTGATGTCTGGC 13814494 14513 TCACAGCTCACCGAGTCTGC 98 14524 14543 AGTGTAAAATAAAGCCCCTA 9614601 14620 AGGACCCAAGTCATCCTGCT 124 14631 14650 GGCCATCAGCTGGCAATGCT 8214670 14689 AAGGAAAGGGAGGCCTAGAG 133 14675 14694 TAGACAAGGAAAGGGAGGCC103 14681 14700 ATTTCATAGACAAGGAAAGG 155 14801 14820CTTATAGTTAACACACAGAA 156 14809 14828 AAGTCAACCTTATAGTTAAC 215 1487714896 ATACACCTCCACCAGGCTGC 216 14888 14907 GTGTCTAGGAGATACACCTC 19 1489114910 CTGGTGTCTAGGAGATACAC 217 14893 14912 TGCTGGTGTCTAGGAGATAC 20 1489614915 GTATGCTGGTGTCTAGGAGA 21 14916 14935 GATTTCCCGGTGGTCACTCT 218 1491814937 TCGATTTCCCGGTGGTCACT 219 14919 14938 CTCGATTTCCCGGTGGTCAC 22014920 14939 CCTCGATTTCCCGGTGGTCA 221 14921 14940 CCCTCGATTTCCCGGTGGTC 2214922 14941 GCCCTCGATTTCCCGGTGGT 222 14923 14942 TGCCCTCGATTTCCCGGTGG223 14924 14943 CTGCCCTCGATTTCCCGGTG 224 14925 14944CCTGCCCTCGATTTCCCGGT 225 14926 14945 CCCTGCCCTCGATTTCCCGG 226 1493014949 ATGACCCTGCCCTCGATTTC 227 14932 14951 CCATGACCCTGCCCTCGATT 22814934 14953 GACCATGACCCTGCCCTCGA 23 14936 14955 GTGACCATGACCCTGCCCTC 22914938 14957 CGGTGACCATGACCCTGCCC 230 14940 14959 GTCGGTGACCATGACCCTGC231 14942 14961 AAGTCGGTGACCATGACCCT 232 14944 14963CGAAGTCGGTGACCATGACC 24 14946 14965 CTCGAAGTCGGTGACCATGA 233 14954 14973GGCACATTCTCGAAGTCGGT 25 14979 14998 GTGGAAGCGGGTCCCGTCCT 235 15254 15273GGTGGGTGCCATGACTGTCA 236 15257 15276 CCAGGTGGGTGCCATGACTG 237 1526115280 CCTGCCAGGTGGGTGCCATG 26 15264 15283 ACCCCTGCCAGGTGGGTGCC 238 1526615285 CCACCCCTGCCAGGTGGGTG 27 15269 15288 TGACCACCCCTGCCAGGTGG 239 1527115290 GCTGACCACCCCTGCCAGGT 240 15279 15298 TCCCGGCCGCTGACCACCCC 24115283 15302 GGCATCCCGGCCGCTGACCA 242 15286 15305 GCCGGCATCCCGGCCGCTGA243 15291 15310 GCCACGCCGGCATCCCGGCC 244 15292 15311GGCCACGCCGGCATCCCGGC 245 15293 15312 TGGCCACGCCGGCATCCCGG 246 1529415313 TTGGCCACGCCGGCATCCCG 247 15295 15314 CTTGGCCACGCCGGCATCCC 24815296 15315 CCTTGGCCACGCCGGCATCC 249 15297 15316 CCCTTGGCCACGCCGGCATC 2815298 15317 ACCCTTGGCCACGCCGGCAT 447 15298 15317 ACCCTTGGTCACGCCGGCAT250 15299 15318 CACCCTTGGCCACGCCGGCA 251 15300 15319GCACCCTTGGCCACGCCGGC 252 15301 15320 GGCACCCTTGGCCACGCCGG 253 1530215321 TGGCACCCTTGGCCACGCCG 254 15303 15322 CTGGCACCCTTGGCCACGCC 25515304 15323 GCTGGCACCCTTGGCCACGC 256 15309 15328 CGCATGCTGGCACCCTTGGC257 15330 15349 CAGTTGAGCACGCGCAGGCT 258 15332 15351GGCAGTTGAGCACGCGCAGG 29 15334 15353 TTGGCAGTTGAGCACGCGCA 259 15336 15355CCTTGGCAGTTGAGCACGCG 30 15339 15358 TTCCCTTGGCAGTTGAGCAC 260 15341 15360CCTTCCCTTGGCAGTTGAGC 261 15345 15364 GTGCCCTTCCCTTGGCAGTT 262 1534715366 CCGTGCCCTTCCCTTGGCAG 263 15358 15377 GGTGCCGCTAACCGTGCCCT 86 1547115490 ACAGCATTCTTGGTTAGGAG 97 16134 16153 AGTCAAGCTGCTGCCCAGAG 120 1666816687 GCTAGTTATTAAGCACCTGC 150 17267 17286 TGTGAGCTCTGGCCCAGTGG 11518377 18396 GAGTAAGGCAGGTTACTCTC 134 18408 18427 TAGATGTGACTAACATTTAA157 18561 18580 AGGAACAAAGCCAAGGTCAC 266 18591 18610TGGCTTTTCCGAATAAACTC 32 18593 18612 GCTGGCTTTTCCGAATAAAC 267 18595 18614CAGCTGGCTTTTCCGAATAA 268 18597 18616 ACCAGCTGGCTTTTCCGAAT 269 1859918618 GGACCAGCTGGCTTTTCCGA 270 18603 18622 GGCTGGACCAGCTGGCTTTT 27118614 18633 GTGGCCCCACAGGCTGGACC 272 18627 18646 AGCAGCACCACCAGTGGCCC273 18649 18668 GCTGTACCCACCCGCCAGGG 274 18695 18714CGACCCCAGCCCTCGCCAGG 33 18705 18724 GTGACCAGCACGACCCCAGC 275 18707 18726CGGTGACCAGCACGACCCCA 276 18709 18728 AGCGGTGACCAGCACGACCC 277 1871118730 GCAGCGGTGACCAGCACGAC 278 18713 18732 CGGCAGCGGTGACCAGCACG 27918714 18733 CCGGCAGCGGTGACCAGCAC 280 18717 18736 TTGCCGGCAGCGGTGACCAG281 18719 18738 AGTTGCCGGCAGCGGTGACC 282 18721 18740GAAGTTGCCGGCAGCGGTGA 283 18723 18742 CGGAAGTTGCCGGCAGCGGT 284 1872518744 CCCGGAAGTTGCCGGCAGCG 285 18727 18746 GTCCCGGAAGTTGCCGGCAG 10519203 19222 CACATTAGCCTTGCTCAAGT 151 19913 19932 TGTGATGACCTGGAAAGGTG288 19931 19950 GGCATTGGTGGCCCCAACTG 149 19933 19952TGGGCATTGGTGGCCCCAAC 289 19941 19960 GCTGGTCTTGGGCATTGGTG 290 1995419973 CCCAGGGTCACCGGCTGGTC 291 19956 19975 TCCCCAGGGTCACCGGCTGG 29219958 19977 AGTCCCCAGGGTCACCGGCT 293 19960 19979 AAAGTCCCCAGGGTCACCGG 3419962 19981 CCAAAGTCCCCAGGGTCACC 294 19964 19983 CCCCAAAGTCCCCAGGGTCA128 19966 19985 GTCCCCAAAGTCCCCAGGGT 295 19969 19988TTGGTCCCCAAAGTCCCCAG 35 19971 19990 AGTTGGTCCCCAAAGTCCCC 296 19973 19992AAAGTTGGTCCCCAAAGTCC 36 19976 19995 GCCAAAGTTGGTCCCCAAAG 297 19978 19997CGGCCAAAGTTGGTCCCCAA 298 19980 19999 AGCGGCCAAAGTTGGTCCCC 299 1998220001 ACAGCGGCCAAAGTTGGTCC 300 19984 20003 ACACAGCGGCCAAAGTTGGT 30119986 20005 CCACACAGCGGCCAAAGTTG 37 19988 20007 GTCCACACAGCGGCCAAAGT 30219990 20009 AGGTCCACACAGCGGCCAAA 303 19992 20011 AGAGGTCCACACAGCGGCCA304 19994 20013 AAAGAGGTCCACACAGCGGC 38 19997 20016 GGCAAAGAGGTCCACACAGC305 20016 20035 CAATGATGTCCTCCCCTGGG 306 20023 20042GAGGCACCAATGATGTCCTC 39 20025 20044 TGGAGGCACCAATGATGTCC 307 20027 20046GCTGGAGGCACCAATGATGT 308 20029 20048 TCGCTGGAGGCACCAATGAT 309 2003120050 AGTCGCTGGAGGCACCAATG 310 20033 20052 GCAGTCGCTGGAGGCACCAA 40 2003620055 GCTGCAGTCGCTGGAGGCAC 311 20038 20057 GTGCTGCAGTCGCTGGAGGC 31220040 20059 AGGTGCTGCAGTCGCTGGAG 313 20042 20061 GCAGGTGCTGCAGTCGCTGG314 20043 20062 AGCAGGTGCTGCAGTCGCTG 315 20045 20064AAAGCAGGTGCTGCAGTCGC 41 20047 20066 ACAAAGCAGGTGCTGCAGTC 316 20049 20068ACACAAAGCAGGTGCTGCAG 317 20051 20070 TGACACAAAGCAGGTGCTGC 318 2006120080 TCCCACTCTGTGACACAAAG 158 20100 20119 GTGGTGACTTACCAGCCACG 10920188 20207 CCCCTGCACAGAGCCTGGCA 141 20624 20643 TCATGGCTGCAATGCCTGGT320 20629 20648 CAGCATCATGGCTGCAATGC 321 20631 20650GACAGCATCATGGCTGCAAT 322 20633 20652 CAGACAGCATCATGGCTGCA 43 20635 20654GGCAGACAGCATCATGGCTG 323 20637 20656 TCGGCAGACAGCATCATGGC 324 2063920658 GCTCGGCAGACAGCATCATG 325 20641 20660 CGGCTCGGCAGACAGCATCA 32620643 20662 TCCGGCTCGGCAGACAGCAT 327 20657 20676 CGGCCAGGGTGAGCTCCGGC328 20670 20689 CTCTGCCTCAACTCGGCCAG 329 20672 20691GTCTCTGCCTCAACTCGGCC 330 20674 20693 CAGTCTCTGCCTCAACTCGG 331 2067620695 ATCAGTCTCTGCCTCAACTC 332 20678 20697 GGATCAGTCTCTGCCTCAAC 33320680 20699 GTGGATCAGTCTCTGCCTCA 334 20682 20701 AAGTGGATCAGTCTCTGCCT 4420683 20702 GAAGTGGATCAGTCTCTGCC 335 20685 20704 GAGAAGTGGATCAGTCTCTG336 20687 20706 CAGAGAAGTGGATCAGTCTC 337 20689 20708GGCAGAGAAGTGGATCAGTC 45 20691 20710 TTGGCAGAGAAGTGGATCAG 338 20693 20712CTTTGGCAGAGAAGTGGATC 46 20696 20715 CATCTTTGGCAGAGAAGTGG 339 20698 20717GACATCTTTGGCAGAGAAGT 340 20700 20719 ATGACATCTTTGGCAGAGAA 47 20702 20721TGATGACATCTTTGGCAGAG 341 20704 20723 ATTGATGACATCTTTGGCAG 342 2070620725 TCATTGATGACATCTTTGGC 48 20709 20728 GCCTCATTGATGACATCTTT 343 2071120730 AGGCCTCATTGATGACATCT 344 20713 20732 CCAGGCCTCATTGATGACAT 34520715 20734 AACCAGGCCTCATTGATGAC 346 20717 20736 GGAACCAGGCCTCATTGATG347 20718 20737 GGGAACCAGGCCTCATTGAT 348 20719 20738AGGGAACCAGGCCTCATTGA 349 20720 20739 CAGGGAACCAGGCCTCATTG 49 20721 20740TCAGGGAACCAGGCCTCATT 350 20722 20741 CTCAGGGAACCAGGCCTCAT 351 2072320742 CCTCAGGGAACCAGGCCTCA 352 20724 20743 TCCTCAGGGAACCAGGCCTC 35320725 20744 GTCCTCAGGGAACCAGGCCT 50 20726 20745 GGTCCTCAGGGAACCAGGCC 35420727 20746 TGGTCCTCAGGGAACCAGGC 355 20728 20747 CTGGTCCTCAGGGAACCAGG356 20729 20748 GCTGGTCCTCAGGGAACCAG 357 20730 20749CGCTGGTCCTCAGGGAACCA 87 20733 20752 ACCCGCTGGTCCTCAGGGAA 358 20735 20754GTACCCGCTGGTCCTCAGGG 359 20737 20756 CAGTACCCGCTGGTCCTCAG 51 20740 20759GGTCAGTACCCGCTGGTCCT 119 20762 20781 GCAGGGCGGCCACCAGGTTG 360 2078520804 ACCTGCCCCATGGGTGCTGG 93 20995 21014 AGAGAGGAGGGCTTAAAGAA 95 2108221101 AGCTGCCAACCTGCAAAAAG 361 21088 21107 CAAAACAGCTGCCAACCTGC 53 2109121110 CTGCAAAACAGCTGCCAACC 362 21093 21112 TCCTGCAAAACAGCTGCCAA 36321095 21114 AGTCCTGCAAAACAGCTGCC 364 21106 21125 GCTGACCATACAGTCCTGCA365 21118 21137 GGCCCCGAGTGTGCTGACCA 54 21121 21140 GTAGGCCCCGAGTGTGCTGA366 21123 21142 GTGTAGGCCCCGAGTGTGCT 367 21125 21144CCGTGTAGGCCCCGAGTGTG 368 21127 21146 ATCCGTGTAGGCCCCGAGTG 369 2112921148 CCATCCGTGTAGGCCCCGAG 370 21131 21150 GGCCATCCGTGTAGGCCCCG 37121133 21152 GTGGCCATCCGTGTAGGCCC 372 21181 21200 CTGGAGCAGCTCAGCAGCTC460 21181 21194 CAGCTCAGCAGCTC 373 21183 21202 AACTGGAGCAGCTCAGCAGC 5521186 21205 AGAAACTGGAGCAGCTCAGC 374 21188 21207 GGAGAAACTGGAGCAGCTCA375 21190 21209 CTGGAGAAACTGGAGCAGCT 56 21192 21211 TCCTGGAGAAACTGGAGCAG143 21481 21500 TGAAAATCCATCCAGCACTG 89 21589 21608 AGAACCATGGAGCACCTGAG448 21692 21711 CTGCCCTTCCACCAAAATGC 449 21693 21712ACTGCCCTTCCACCAAAATG 450 21694 21713 CACTGCCCTTCCACCAAAAT 451 2169521714 GCACTGCCCTTCCACCAAAA 123 21696 21715 GGCACTGCCCTTCCACCAAA 45221697 21716 GGGCACTGCCCTTCCACCAA 453 21698 21717 TGGGCACTGCCCTTCCACCA454 21699 21718 CTGGGCACTGCCCTTCCACC 455 21700 21719GCTGGGCACTGCCCTTCCAC 57 22096 22115 CGTTGTGGGCCCGGCAGACC 376 22133 22152CACCTGGCAATGGCGTAGAC 377 22134 22153 GCACCTGGCAATGGCGTAGA 378 2213522154 AGCACCTGGCAATGGCGTAG 379 22136 22155 CAGCACCTGGCAATGGCGTA 38022137 22156 GCAGCACCTGGCAATGGCGT 381 22138 22157 GGCAGCACCTGGCAATGGCG382 22139 22158 AGGCAGCACCTGGCAATGGC 383 22140 22159CAGGCAGCACCTGGCAATGG 384 22141 22160 GCAGGCAGCACCTGGCAATG 58 22142 22161AGCAGGCAGCACCTGGCAAT 385 22143 22162 TAGCAGGCAGCACCTGGCAA 386 2214422163 GTAGCAGGCAGCACCTGGCA 387 22189 22208 TGGCCTCAGCTGGTGGAGCT 38822199 22218 GTCCCCATGCTGGCCTCAGC 389 22200 22219 GGTCCCCATGCTGGCCTCAG390 22201 22220 GGGTCCCCATGCTGGCCTCA 391 22202 22221CGGGTCCCCATGCTGGCCTC 392 22203 22222 ACGGGTCCCCATGCTGGCCT 59 22204 22223CACGGGTCCCCATGCTGGCC 393 22205 22224 ACACGGGTCCCCATGCTGGC 394 2220622225 GACACGGGTCCCCATGCTGG 395 22207 22226 GGACACGGGTCCCCATGCTG 39622208 22227 TGGACACGGGTCCCCATGCT 397 22209 22228 GTGGACACGGGTCCCCATGC398 22210 22229 AGTGGACACGGGTCCCCATG 399 22211 22230CAGTGGACACGGGTCCCCAT 400 22212 22231 GCAGTGGACACGGGTCCCCA 401 2221322232 GGCAGTGGACACGGGTCCCC 402 22214 22233 TGGCAGTGGACACGGGTCCC 40322215 22234 GTGGCAGTGGACACGGGTCC 404 22216 22235 GGTGGCAGTGGACACGGGTC405 22217 22236 TGGTGGCAGTGGACACGGGT 406 22220 22239TGTTGGTGGCAGTGGACACG 126 22292 22311 GGTGCATAAGGAGAAAGAGA 408 2398524004 GTGCCAAGGTCCTCCACCTC 409 24005 24024 TCAGCACAGGCGGCTTGTGG 41024035 24054 CCACGCACTGGTTGGGCTGA 60 24095 24114 CTTTGCATTCCAGACCTGGG 41124096 24115 ACTTTGCATTCCAGACCTGG 412 24097 24116 GACTTTGCATTCCAGACCTG413 24098 24117 TGACTTTGCATTCCAGACCT 414 24099 24118TTGACTTTGCATTCCAGACC 61 24100 24119 CTTGACTTTGCATTCCAGAC 415 24102 24121TCCTTGACTTTGCATTCCAG 416 24115 24134 CGGGATTCCATGCTCCTTGA 147 2485824877 TGAGTTCATTTAAGAGTGGA 118 24907 24926 GCACCATCCAGACCAGAATC 11425413 25432 GAGAGGTTCAGATCCAGGCC 418 25994 26013 GGAGGGCACTGCAGCCAGTC419 26112 26131 CAGATGGCAACGGCTGTCAC 420 26113 26132GCAGATGGCAACGGCTGTCA 421 26114 26133 AGCAGATGGCAACGGCTGTC 422 2611526134 CAGCAGATGGCAACGGCTGT 423 26116 26135 GCAGCAGATGGCAACGGCTG 62 2611726136 GGCAGCAGATGGCAACGGCT 424 26118 26137 CGGCAGCAGATGGCAACGGC 42526119 26138 CCGGCAGCAGATGGCAACGG 426 26120 26139 TCCGGCAGCAGATGGCAACG427 26121 26140 CTCCGGCAGCAGATGGCAAC 428 26122 26141GCTCCGGCAGCAGATGGCAA 429 26132 26151 CCAGGTGCCGGCTCCGGCAG 430 2614226161 GAGGCCTGCGCCAGGTGCCG 154 26217 26236 TTTTAAAGCTCAGCCCCAGC 63 2622226241 AACCATTTTAAAGCTCAGCC 64 26311 26330 TCAAGGGCCAGGCCAGCAGC 65 2631626335 CCCACTCAAGGGCCAGGCCA 122 26389 26408 GGAGGGAGCTTCCTGGCACC 66 2640426423 ATGCCCCACAGTGAGGGAGG 67 26413 26432 AATGGTGAAATGCCCCACAG 153 2645626475 TTGGGAGCAGCTGGCAGCAC 68 26557 26576 CATGGGAAGAATCCTGCCTC 431 2663526654 ATGAGGGCCATCAGCACCTT 432 26636 26655 GATGAGGGCCATCAGCACCT 43326637 26656 AGATGAGGGCCATCAGCACC 434 26638 26657 GAGATGAGGGCCATCAGCAC 6926639 26658 GGAGATGAGGGCCATCAGCA 435 26640 26659 TGGAGATGAGGGCCATCAGC436 26641 26660 CTGGAGATGAGGGCCATCAG 437 26642 26661GCTGGAGATGAGGGCCATCA 438 26643 26662 AGCTGGAGATGAGGGCCATC 461 2668426697 TTAATCAGGGAGCC 70 26707 26726 TAGATGCCATCCAGAAAGCT 439 26709 26728GCTAGATGCCATCCAGAAAG 440 26710 26729 GGCTAGATGCCATCCAGAAA 441 2671126730 TGGCTAGATGCCATCCAGAA 442 26712 26731 CTGGCTAGATGCCATCCAGA 71 2671326732 TCTGGCTAGATGCCATCCAG 443 26714 26733 CTCTGGCTAGATGCCATCCA 44426715 26734 CCTCTGGCTAGATGCCATCC 445 26716 26735 GCCTCTGGCTAGATGCCATC446 26717 26736 AGCCTCTGGCTAGATGCCAT 72 26790 26809 GGCATAGAGCAGAGTAAAGG73 26795 26814 AGCCTGGCATAGAGCAGAGT 135 26801 26820 TAGCACAGCCTGGCATAGAG112 27034 27053 GAAGAGGCTTGGCTTCAGAG 74 27040 27059 AAGTAAGAAGAGGCTTGGCT75 27244 27263 GCTCAAGGAGGGACAGTTGT 76 27279 27298 AAAGATAAATGTCTGCTTGC77 27284 27303 ACCCAAAAGATAAATGTCTG 78 27350 27369 TCTTCAAGTTACAAAAGCAA99 27357 27376 ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to aPCSK9 nucleic acid. In certain such embodiments, gapmer antisensecompounds are targeted to SEQ ID NO: 2. In certain such embodiments, thenucleotide sequences illustrated in Table 7 have a 5-10-5 gapmer motif.Table 8 illustrates gapmer antisense compounds targeted to SEQ ID NO: 2,having a 5-10-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 8 Gapmer antisense compounds having a 5-10-5 motif targeted to SEQID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID IsisNo Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 395149 5-10-5 4 22742293 0 GCGCGGAATCCTGGCTGGGA 395150 5-10-5 5 2381 2400 0GAGGAGACCTAGAGGCCGTG 395151 5-10-5 6 2439 2458 0 AGGACCGCCTGGAGCTGACG395152 5-10-5 7 2549 2568 0 ACGCAAGGCTAGCACCAGCT 399793 5-10-5 8 25562575 0 CCTCGGAACGCAAGGCTAGC 395153 5-10-5 9 2619 2638 0CCTTGGCGCAGCGGTGGAAG 399837 5-10-5 107 3056 3075 0 CCCACTATAATGGCAAGCCC399838 5-10-5 80 4306 4325 0 AACCCAGTTCTAATGCACCT 399839 5-10-5 106 51405159 0 CCAGTCAGAGTAGAACAGAG 395221 5-10-5 102 5590 5609 0ATGTGCAGAGATCAATCACA 399840 5-10-5 121 5599 5618 0 GGAGCCTACATGTGCAGAGA399841 5-10-5 94 5667 5686 0 AGCATGGCACCAGCATCTGC 395154 5-10-5 10 64986517 0 GGCGGGCAGTGCGCTCTGAC 395155 5-10-5 11 6537 6556 0TGGTGAGGTATCCCCGGCGG 399794 5-10-5 12 6543 6562 0 GGATCTTGGTGAGGTATCCC399795 5-10-5 13 6552 6571 0 AGACATGCAGGATCTTGGTG 395156 5-10-5 14 65576576 0 ATGGAAGACATGCAGGATCT 395157 5-10-5 15 6583 6602 0TTCACCAGGAAGCCAGGAAG 399796 5-10-5 16 6588 6607 0 TCATCTTCACCAGGAAGCCA399842 5-10-5 108 6652 6671 0 CCCAGCCCTATCAGGAAGTG 399843 5-10-5 1447099 7118 0 TGACATCCAGGAGGGAGGAG 399844 5-10-5 91 7556 7575 0AGACTGATGGAAGGCATTGA 399845 5-10-5 131 7565 7584 0 GTGTTGAGCAGACTGATGGA399846 5-10-5 145 8836 8855 0 TGACATCTTGTCTGGGAGCC 399847 5-10-5 90 89488967 0 AGACTAGGAGCCTGAGTTTT 399848 5-10-5 125 9099 9118 0GGCCTGCAGAAGCCAGAGAG 395158 5-10-5 17 9130 9149 0 CCTCGATGTAGTCGACATGG395159 5-10-5 18 9210 9229 0 GTATTCATCCGCCCGGTACC 399849 5-10-5 14810252 10271 0 TGGCAGCAACTCAGACATAT 395222 5-10-5 127 10633 10652 0GGTGGTAATTTGTCACAGCA 395223 5-10-5 84 11308 11327 0 AAGGTCACACAGTTAAGAGT399850 5-10-5 79 11472 11491 0 AAATGCAGGGCTAAAATCAC 399851 5-10-5 8812715 12734 0 ACTGGATACATTGGCAGACA 399852 5-10-5 111 12928 12947 0CTAGAGGAACCACTAGATAT 395201 5-10-5 85 13681 13700 0 ACAAATTCCCAGACTCAGCA399827 5-10-5 100 13746 13765 0 ATCTCAGGACAGGTGAGCAA 399828 5-10-5 11613760 13779 0 GAGTAGAGATTCTCATCTCA 395202 5-10-5 129 13816 13835 0GTGCCATCTGAACAGCACCT 399829 5-10-5 117 13828 13847 0GAGTCTTCTGAAGTGCCATC 399830 5-10-5 81 13903 13922 0 AAGCAGGGCCTCAGGTGGAA395203 5-10-5 110 13926 13945 0 CCTGGAACCCCTGCAGCCAG 395204 5-10-5 15213977 13996 0 TTCAGGCAGGTTGCTGCTAG 399831 5-10-5 83 13986 14005 0AAGGAAGACTTCAGGCAGGT 395205 5-10-5 140 13998 14017 0TCAGCCAGGCCAAAGGAAGA 399832 5-10-5 137 14112 14131 0TAGGGAGAGCTCACAGATGC 395206 5-10-5 136 14122 14141 0TAGGAGAAAGTAGGGAGAGC 395207 5-10-5 132 14179 14198 0TAAAAGCTGCAAGAGACTCA 395208 5-10-5 139 14267 14286 0TCAGAGAAAACAGTCACCGA 399833 5-10-5 92 14397 14416 0 AGAGACAGGAAGCTGCAGCT395209 5-10-5 142 14404 14423 0 TCATTTTAGAGACAGGAAGC 395210 5-10-5 11314441 14460 0 GAATAACAGTGATGTCTGGC 395211 5-10-5 138 14494 14513 0TCACAGCTCACCGAGTCTGC 395212 5-10-5 98 14524 14543 0 AGTGTAAAATAAAGCCCCTA395213 5-10-5 96 14601 14620 0 AGGACCCAAGTCATCCTGCT 395214 5-10-5 12414631 14650 0 GGCCATCAGCTGGCAATGCT 399834 5-10-5 82 14670 14689 0AAGGAAAGGGAGGCCTAGAG 395215 5-10-5 133 14675 14694 0TAGACAAGGAAAGGGAGGCC 395216 5-10-5 103 14681 14700 0ATTTCATAGACAAGGAAAGG 395217 5-10-5 155 14801 14820 0CTTATAGTTAACACACAGAA 399835 5-10-5 156 14809 14828 0AAGTCAACCTTATAGTTAAC 395160 5-10-5 19 14891 14910 0 CTGGTGTCTAGGAGATACAC399797 5-10-5 20 14896 14915 0 GTATGCTGGTGTCTAGGAGA 395161 5-10-5 2114916 14935 0 GATTTCCCGGTGGTCACTCT 399798 5-10-5 22 14922 14941 0GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 14936 14955 0 GTGACCATGACCCTGCCCTC395162 5-10-5 24 14946 14965 0 CTCGAAGTCGGTGACCATGA 395163 5-10-5 2514979 14998 0 GTGGAAGCGGGTCCCGTCCT 395164 5-10-5 26 15264 15283 0ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 15269 15288 0 TGACCACCCCTGCCAGGTGG395165 5-10-5 28 15298 15317 0 ACCCTTGGCCACGCCGGCAT 395166 5-10-5 2915334 15353 0 TTGGCAGTTGAGCACGCGCA 399801 5-10-5 30 15339 15358 0TTCCCTTGGCAGTTGAGCAC 399853 5-10-5 86 15471 15490 0 ACAGCATTCTTGGTTAGGAG399854 5-10-5 97 16134 16153 0 AGTCAAGCTGCTGCCCAGAG 399855 5-10-5 12016668 16687 0 GCTAGTTATTAAGCACCTGC 399856 5-10-5 150 17267 17286 0TGTGAGCTCTGGCCCAGTGG 399857 5-10-5 115 18377 18396 0GAGTAAGGCAGGTTACTCTC 399858 5-10-5 134 18408 18427 0TAGATGTGACTAACATTTAA 395224 5-10-5 157 18561 18580 0AGGAACAAAGCCAAGGTCAC 395168 5-10-5 32 18593 18612 0 GCTGGCTTTTCCGAATAAAC395169 5-10-5 33 18705 18724 0 GTGACCAGCACGACCCCAGC 399859 5-10-5 10519203 19222 0 CACATTAGCCTTGCTCAAGT 399860 5-10-5 151 19913 19932 0TGTGATGACCTGGAAAGGTG 395170 5-10-5 149 19933 19952 0TGGGCATTGGTGGCCCCAAC 395171 5-10-5 34 19962 19981 0 CCAAAGTCCCCAGGGTCACC395172 5-10-5 128 19966 19985 0 GTCCCCAAAGTCCCCAGGGT 399802 5-10-5 3519971 19990 0 AGTTGGTCCCCAAAGTCCCC 395173 5-10-5 36 19976 19995 0GCCAAAGTTGGTCCCCAAAG 399803 5-10-5 37 19988 20007 0 GTCCACACAGCGGCCAAAGT395174 5-10-5 38 19997 20016 0 GGCAAAGAGGTCCACACAGC 395175 5-10-5 3920025 20044 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 20036 20055 0GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 20047 20066 0 ACAAAGCAGGTGCTGCAGTC399861 5-10-5 158 20100 20119 0 GTGGTGACTTACCAGCCACG 399862 5-10-5 10920188 20207 0 CCCCTGCACAGAGCCTGGCA 399863 5-10-5 141 20624 20643 0TCATGGCTGCAATGCCTGGT 399807 5-10-5 43 20635 20654 0 GGCAGACAGCATCATGGCTG399808 5-10-5 44 20683 20702 0 GAAGTGGATCAGTCTCTGCC 395177 5-10-5 4520691 20710 0 TTGGCAGAGAAGTGGATCAG 399809 5-10-5 46 20696 20715 0CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 20702 20721 0 TGATGACATCTTTGGCAGAG399811 5-10-5 48 20709 20728 0 GCCTCATTGATGACATCTTT 399812 5-10-5 4920721 20740 0 TCAGGGAACCAGGCCTCATT 395178 5-10-5 50 20726 20745 0GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 20733 20752 0 ACCCGCTGGTCCTCAGGGAA399813 5-10-5 51 20740 20759 0 GGTCAGTACCCGCTGGTCCT 395180 5-10-5 11920762 20781 0 GCAGGGCGGCCACCAGGTTG 399864 5-10-5 93 20995 21014 0AGAGAGGAGGGCTTAAAGAA 399865 5-10-5 95 21082 21101 0 AGCTGCCAACCTGCAAAAAG399814 5-10-5 53 21091 21110 0 CTGCAAAACAGCTGCCAACC 395182 5-10-5 5421121 21140 0 GTAGGCCCCGAGTGTGCTGA 399815 5-10-5 55 21186 21205 0AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 21192 21211 0 TCCTGGAGAAACTGGAGCAG399866 5-10-5 143 21481 21500 0 TGAAAATCCATCCAGCACTG 399867 5-10-5 8921589 21608 0 AGAACCATGGAGCACCTGAG 399868 5-10-5 123 21696 21715 0GGCACTGCCCTTCCACCAAA 395183 5-10-5 57 22096 22115 0 CGTTGTGGGCCCGGCAGACC395184 5-10-5 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT 395185 5-10-5 5922204 22223 0 CACGGGTCCCCATGCTGGCC 395225 5-10-5 126 22292 22311 0GGTGCATAAGGAGAAAGAGA 395186 5-10-5 60 24095 24114 0 CTTTGCATTCCAGACCTGGG399817 5-10-5 61 24100 24119 0 CTTGACTTTGCATTCCAGAC 395226 5-10-5 14724858 24877 0 TGAGTTCATTTAAGAGTGGA 399869 5-10-5 118 24907 24926 0GCACCATCCAGACCAGAATC 399870 5-10-5 114 25413 25432 0GAGAGGTTCAGATCCAGGCC 395187 5-10-5 62 26117 26136 0 GGCAGCAGATGGCAACGGCT395188 5-10-5 154 26217 26236 0 TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 6326222 26241 0 AACCATTTTAAAGCTCAGCC 395189 5-10-5 64 26311 26330 0TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 26316 26335 0 CCCACTCAAGGGCCAGGCCA399820 5-10-5 122 26389 26408 0 GGAGGGAGCTTCCTGGCACC 395190 5-10-5 6626404 26423 0 ATGCCCCACAGTGAGGGAGG 395191 5-10-5 67 26413 26432 0AATGGTGAAATGCCCCACAG 395192 5-10-5 153 26456 26475 0TTGGGAGCAGCTGGCAGCAC 395193 5-10-5 68 26557 26576 0 CATGGGAAGAATCCTGCCTC395194 5-10-5 69 26639 26658 0 GGAGATGAGGGCCATCAGCA 395195 5-10-5 7026707 26726 0 TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 26713 26732 0TCTGGCTAGATGCCATCCAG 395196 5-10-5 72 26790 26809 0 GGCATAGAGCAGAGTAAAGG399822 5-10-5 73 26795 26814 0 AGCCTGGCATAGAGCAGAGT 399823 5-10-5 13526801 26820 0 TAGCACAGCCTGGCATAGAG 395197 5-10-5 112 27034 27053 0GAAGAGGCTTGGCTTCAGAG 399824 5-10-5 74 27040 27059 0 AAGTAAGAAGAGGCTTGGCT395198 5-10-5 75 27244 27263 0 GCTCAAGGAGGGACAGTTGT 395199 5-10-5 7627279 27298 0 AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 27284 27303 0ACCCAAAAGATAAATGTCTG 395200 5-10-5 78 27350 27369 0 TCTTCAAGTTACAAAAGCAA399826 5-10-5 99 27357 27376 0 ATAAATATCTTCAAGTTACA 405861 5-10-5 1622545 2564 0 AAGGCTAGCACCAGCTCCTC 405862 5-10-5 163 2546 2565 0CAAGGCTAGCACCAGCTCCT 405863 5-10-5 164 2547 2566 0 GCAAGGCTAGCACCAGCTCC405864 5-10-5 165 2548 2567 0 CGCAAGGCTAGCACCAGCTC 405865 5-10-5 1662550 2569 0 AACGCAAGGCTAGCACCAGC 405866 5-10-5 167 2551 2570 0GAACGCAAGGCTAGCACCAG 405867 5-10-5 168 2552 2571 0 GGAACGCAAGGCTAGCACCA405868 5-10-5 169 2553 2572 0 CGGAACGCAAGGCTAGCACC 405869 5-10-5 21814918 14937 0 TCGATTTCCCGGTGGTCACT 405870 5-10-5 219 14919 14938 0CTCGATTTCCCGGTGGTCAC 405871 5-10-5 220 14920 14939 0CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 14921 14940 0CCCTCGATTTCCCGGTGGTC 405873 5-10-5 222 14923 14942 0TGCCCTCGATTTCCCGGTGG 405874 5-10-5 223 14924 14943 0CTGCCCTCGATTTCCCGGTG 405875 5-10-5 224 14925 14944 0CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 14926 14945 0CCCTGCCCTCGATTTCCCGG 405877 5-10-5 246 15294 15313 0TTGGCCACGCCGGCATCCCG 405878 5-10-5 247 15295 15314 0CTTGGCCACGCCGGCATCCC 405879 5-10-5 248 15296 15315 0CCTTGGCCACGCCGGCATCC 405880 5-10-5 249 15297 15316 0CCCTTGGCCACGCCGGCATC 405881 5-10-5 250 15299 15318 0CACCCTTGGCCACGCCGGCA 405882 5-10-5 251 15300 15319 0GCACCCTTGGCCACGCCGGC 405883 5-10-5 252 15301 15320 0GGCACCCTTGGCCACGCCGG 405884 5-10-5 253 15302 15321 0TGGCACCCTTGGCCACGCCG 405885 5-10-5 346 20717 20736 0GGAACCAGGCCTCATTGATG 405886 5-10-5 347 20718 20737 0GGGAACCAGGCCTCATTGAT 405887 5-10-5 348 20719 20738 0AGGGAACCAGGCCTCATTGA 405888 5-10-5 349 20720 20739 0CAGGGAACCAGGCCTCATTG 405889 5-10-5 350 20722 20741 0CTCAGGGAACCAGGCCTCAT 405890 5-10-5 351 20723 20742 0CCTCAGGGAACCAGGCCTCA 405891 5-10-5 352 20724 20743 0TCCTCAGGGAACCAGGCCTC 405892 5-10-5 353 20725 20744 0GTCCTCAGGGAACCAGGCCT 405893 5-10-5 431 26635 26654 0ATGAGGGCCATCAGCACCTT 405894 5-10-5 432 26636 26655 0GATGAGGGCCATCAGCACCT 405895 5-10-5 433 26637 26656 0AGATGAGGGCCATCAGCACC 405896 5-10-5 434 26638 26657 0GAGATGAGGGCCATCAGCAC 405897 5-10-5 435 26640 26659 0TGGAGATGAGGGCCATCAGC 405898 5-10-5 436 26641 26660 0CTGGAGATGAGGGCCATCAG 405899 5-10-5 437 26642 26661 0GCTGGAGATGAGGGCCATCA 405900 5-10-5 438 26643 26662 0AGCTGGAGATGAGGGCCATC 405901 5-10-5 439 26709 26728 0GCTAGATGCCATCCAGAAAG 405902 5-10-5 440 26710 26729 0GGCTAGATGCCATCCAGAAA 405903 5-10-5 441 26711 26730 0TGGCTAGATGCCATCCAGAA 405904 5-10-5 442 26712 26731 0CTGGCTAGATGCCATCCAGA 405905 5-10-5 443 26714 26733 0CTCTGGCTAGATGCCATCCA 405906 5-10-5 444 26715 26734 0CCTCTGGCTAGATGCCATCC 405907 5-10-5 445 26716 26735 0GCCTCTGGCTAGATGCCATC 405908 5-10-5 446 26717 26736 0AGCCTCTGGCTAGATGCCAT 405909 5-10-5 267 18595 18614 0CAGCTGGCTTTTCCGAATAA 405910 5-10-5 268 18597 18616 0ACCAGCTGGCTTTTCCGAAT 405911 5-10-5 269 18599 18618 0GGACCAGCTGGCTTTTCCGA 405912 5-10-5 270 18603 18622 0GGCTGGACCAGCTGGCTTTT 405913 5-10-5 275 18707 18726 0CGGTGACCAGCACGACCCCA 405914 5-10-5 276 18709 18728 0AGCGGTGACCAGCACGACCC 405915 5-10-5 277 18711 18730 0GCAGCGGTGACCAGCACGAC 405916 5-10-5 278 18713 18732 0CGGCAGCGGTGACCAGCACG 405917 5-10-5 280 18717 18736 0TTGCCGGCAGCGGTGACCAG 405918 5-10-5 281 18719 18738 0AGTTGCCGGCAGCGGTGACC 405919 5-10-5 282 18721 18740 0GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 283 18723 18742 0CGGAAGTTGCCGGCAGCGGT 405921 5-10-5 284 18725 18744 0CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 285 18727 18746 0GTCCCGGAAGTTGCCGGCAG 405923 5-10-5 288 19931 19950 0GGCATTGGTGGCCCCAACTG 405924 5-10-5 290 19954 19973 0CCCAGGGTCACCGGCTGGTC 405925 5-10-5 292 19958 19977 0AGTCCCCAGGGTCACCGGCT 405926 5-10-5 293 19960 19979 0AAAGTCCCCAGGGTCACCGG 405927 5-10-5 294 19964 19983 0CCCCAAAGTCCCCAGGGTCA 405928 5-10-5 295 19969 19988 0TTGGTCCCCAAAGTCCCCAG 405929 5-10-5 296 19973 19992 0AAAGTTGGTCCCCAAAGTCC 405930 5-10-5 297 19978 19997 0CGGCCAAAGTTGGTCCCCAA 405931 5-10-5 298 19980 19999 0AGCGGCCAAAGTTGGTCCCC 405932 5-10-5 299 19982 20001 0ACAGCGGCCAAAGTTGGTCC 405933 5-10-5 300 19984 20003 0ACACAGCGGCCAAAGTTGGT 405934 5-10-5 301 19986 20005 0CCACACAGCGGCCAAAGTTG 405935 5-10-5 302 19990 20009 0AGGTCCACACAGCGGCCAAA 405936 5-10-5 304 19994 20013 0AAAGAGGTCCACACAGCGGC 405937 5-10-5 306 20023 20042 0GAGGCACCAATGATGTCCTC 405938 5-10-5 307 20027 20046 0GCTGGAGGCACCAATGATGT 405939 5-10-5 308 20029 20048 0TCGCTGGAGGCACCAATGAT 405940 5-10-5 309 20031 20050 0AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 20033 20052 0GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 20038 20057 0GTGCTGCAGTCGCTGGAGGC 405943 5-10-5 312 20040 20059 0AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 314 20043 20062 0AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 20045 20064 0AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 20049 20068 0ACACAAAGCAGGTGCTGCAG 405947 5-10-5 317 20051 20070 0TGACACAAAGCAGGTGCTGC 405949 5-10-5 320 20629 20648 0CAGCATCATGGCTGCAATGC 405950 5-10-5 321 20631 20650 0GACAGCATCATGGCTGCAAT 405951 5-10-5 322 20633 20652 0CAGACAGCATCATGGCTGCA 405952 5-10-5 323 20637 20656 0TCGGCAGACAGCATCATGGC 405953 5-10-5 325 20641 20660 0CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 20643 20662 0TCCGGCTCGGCAGACAGCAT 405955 5-10-5 328 20670 20689 0CTCTGCCTCAACTCGGCCAG 405956 5-10-5 329 20672 20691 0GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 20674 20693 0CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 20676 20695 0ATCAGTCTCTGCCTCAACTC 405959 5-10-5 332 20678 20697 0GGATCAGTCTCTGCCTCAAC 405960 5-10-5 333 20680 20699 0GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 20682 20701 0AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 20685 20704 0GAGAAGTGGATCAGTCTCTG 405963 5-10-5 337 20689 20708 0GGCAGAGAAGTGGATCAGTC 405964 5-10-5 338 20693 20712 0CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 20698 20717 0GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 20700 20719 0ATGACATCTTTGGCAGAGAA 405967 5-10-5 341 20704 20723 0ATTGATGACATCTTTGGCAG 405968 5-10-5 342 20706 20725 0TCATTGATGACATCTTTGGC 405969 5-10-5 343 20711 20730 0AGGCCTCATTGATGACATCT 405970 5-10-5 344 20713 20732 0CCAGGCCTCATTGATGACAT 405971 5-10-5 355 20728 20747 0CTGGTCCTCAGGGAACCAGG 405972 5-10-5 357 20730 20749 0CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 20735 20754 0GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 20737 20756 0CAGTACCCGCTGGTCCTCAG 405975 5-10-5 361 21088 21107 0CAAAACAGCTGCCAACCTGC 405976 5-10-5 362 21093 21112 0TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 21095 21114 0AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 21118 21137 0GGCCCCGAGTGTGCTGACCA 405979 5-10-5 366 21123 21142 0GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 367 21125 21144 0CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 21127 21146 0ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 21131 21150 0GGCCATCCGTGTAGGCCCCG 405983 5-10-5 371 21133 21152 0GTGGCCATCCGTGTAGGCCC 405984 5-10-5 372 21181 21200 0CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 21183 21202 0AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 21188 21207 0GGAGAAACTGGAGCAGCTCA 405987 5-10-5 375 21190 21209 0CTGGAGAAACTGGAGCAGCT 405988 5-10-5 381 22138 22157 0GGCAGCACCTGGCAATGGCG 405989 5-10-5 383 22140 22159 0CAGGCAGCACCTGGCAATGG 405990 5-10-5 386 22144 22163 0GTAGCAGGCAGCACCTGGCA 405991 5-10-5 394 22206 22225 0GACACGGGTCCCCATGCTGG 405992 5-10-5 396 22208 22227 0TGGACACGGGTCCCCATGCT 405993 5-10-5 398 22210 22229 0AGTGGACACGGGTCCCCATG 405994 5-10-5 400 22212 22231 0GCAGTGGACACGGGTCCCCA 405995 5-10-5 402 22214 22233 0TGGCAGTGGACACGGGTCCC 405996 5-10-5 412 24097 24116 0GACTTTGCATTCCAGACCTG 405997 5-10-5 415 24102 24121 0TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 26120 26139 0TCCGGCAGCAGATGGCAACG 405999 5-10-5 160 2437 2456 0 GACCGCCTGGAGCTGACGGT406003 5-10-5 178 6492 6511 0 CAGTGCGCTCTGACTGCGAG 406004 5-10-5 1796494 6513 0 GGCAGTGCGCTCTGACTGCG 406005 5-10-5 180 6496 6515 0CGGGCAGTGCGCTCTGACTG 406006 5-10-5 181 6499 6518 0 CGGCGGGCAGTGCGCTCTGA406007 5-10-5 183 6532 6551 0 AGGTATCCCCGGCGGGCAGC 406008 5-10-5 1886539 6558 0 CTTGGTGAGGTATCCCCGGC 406009 5-10-5 190 6541 6560 0ATCTTGGTGAGGTATCCCCG 406010 5-10-5 193 6546 6565 0 GCAGGATCTTGGTGAGGTAT406011 5-10-5 194 6548 6567 0 ATGCAGGATCTTGGTGAGGT 406012 5-10-5 1956550 6569 0 ACATGCAGGATCTTGGTGAG 406013 5-10-5 196 6554 6573 0GAAGACATGCAGGATCTTGG 406014 5-10-5 199 6585 6604 0 TCTTCACCAGGAAGCCAGGA406015 5-10-5 200 6590 6609 0 ACTCATCTTCACCAGGAAGC 406016 5-10-5 2016592 6611 0 CCACTCATCTTCACCAGGAA 406017 5-10-5 203 6596 6615 0GTCGCCACTCATCTTCACCA 406018 5-10-5 204 6598 6617 0 AGGTCGCCACTCATCTTCAC406019 5-10-5 205 6600 6619 0 GCAGGTCGCCACTCATCTTC 406020 5-10-5 2066602 6621 0 CAGCAGGTCGCCACTCATCT 406021 5-10-5 210 9207 9226 0TTCATCCGCCCGGTACCGTG 406022 5-10-5 211 9209 9228 0 TATTCATCCGCCCGGTACCG406023 5-10-5 212 9212 9231 0 TGGTATTCATCCGCCCGGTA 406024 5-10-5 2139214 9233 0 GCTGGTATTCATCCGCCCGG 406025 5-10-5 216 14888 14907 0GTGTCTAGGAGATACACCTC 406026 5-10-5 217 14893 14912 0TGCTGGTGTCTAGGAGATAC 406027 5-10-5 226 14930 14949 0ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 14932 14951 0CCATGACCCTGCCCTCGATT 406029 5-10-5 228 14934 14953 0GACCATGACCCTGCCCTCGA 406030 5-10-5 229 14938 14957 0CGGTGACCATGACCCTGCCC 406031 5-10-5 230 14940 14959 0GTCGGTGACCATGACCCTGC 406032 5-10-5 232 14944 14963 0CGAAGTCGGTGACCATGACC 406033 5-10-5 237 15261 15280 0CCTGCCAGGTGGGTGCCATG 406034 5-10-5 238 15266 15285 0CCACCCCTGCCAGGTGGGTG 406035 5-10-5 239 15271 15290 0GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 15283 15302 0GGCATCCCGGCCGCTGACCA 406037 5-10-5 242 15286 15305 0GCCGGCATCCCGGCCGCTGA 406038 5-10-5 245 15293 15312 0TGGCCACGCCGGCATCCCGG 406039 5-10-5 257 15330 15349 0CAGTTGAGCACGCGCAGGCT 406040 5-10-5 258 15332 15351 0GGCAGTTGAGCACGCGCAGG 406041 5-10-5 259 15336 15355 0CCTTGGCAGTTGAGCACGCG 406042 5-10-5 260 15341 15360 0CCTTCCCTTGGCAGTTGAGC 406043 5-10-5 261 15345 15364 0GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 262 15347 15366 0CCGTGCCCTTCCCTTGGCAG 406045 5-10-5 266 18591 18610 0TGGCTTTTCCGAATAAACTC 406478 5-10-5 448 21692 21711 0CTGCCCTTCCACCAAAATGC 406479 5-10-5 449 21693 21712 0ACTGCCCTTCCACCAAAATG 406480 5-10-5 450 21694 21713 0CACTGCCCTTCCACCAAAAT 406481 5-10-5 451 21695 21714 0GCACTGCCCTTCCACCAAAA 406482 5-10-5 452 21697 21716 0GGGCACTGCCCTTCCACCAA 406483 5-10-5 453 21698 21717 0TGGGCACTGCCCTTCCACCA 406484 5-10-5 454 21699 21718 0CTGGGCACTGCCCTTCCACC 406485 5-10-5 455 21700 21719 0GCTGGGCACTGCCCTTCCAC 408653 5-10-5 354 20727 20746 0TGGTCCTCAGGGAACCAGGC 409126 5-10-5 447 15298 15317 1ACCCTTGGTCACGCCGGCAT 410529 5-10-5 184 6534 6553 0 TGAGGTATCCCCGGCGGGCA410530 5-10-5 185 6535 6554 0 GTGAGGTATCCCCGGCGGGC 410531 5-10-5 1866536 6555 0 GGTGAGGTATCCCCGGCGGG 410532 5-10-5 187 6538 6557 0TTGGTGAGGTATCCCCGGCG 410533 5-10-5 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG410534 5-10-5 191 6542 6561 0 GATCTTGGTGAGGTATCCCC 410535 5-10-5 1926544 6563 0 AGGATCTTGGTGAGGTATCC 410536 5-10-5 243 15291 15310 0GCCACGCCGGCATCCCGGCC 410537 5-10-5 244 15292 15311 0GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 15303 15322 0CTGGCACCCTTGGCCACGCC 410539 5-10-5 255 15304 15323 0GCTGGCACCCTTGGCCACGC 410540 5-10-5 356 20729 20748 0GCTGGTCCTCAGGGAACCAG 410541 5-10-5 376 22133 22152 0CACCTGGCAATGGCGTAGAC 410542 5-10-5 377 22134 22153 0GCACCTGGCAATGGCGTAGA 410543 5-10-5 378 22135 22154 0AGCACCTGGCAATGGCGTAG 410544 5-10-5 379 22136 22155 0CAGCACCTGGCAATGGCGTA 410545 5-10-5 380 22137 22156 0GCAGCACCTGGCAATGGCGT 410546 5-10-5 382 22139 22158 0AGGCAGCACCTGGCAATGGC 410547 5-10-5 384 22141 22160 0GCAGGCAGCACCTGGCAATG 410548 5-10-5 385 22143 22162 0TAGCAGGCAGCACCTGGCAA 410549 5-10-5 388 22199 22218 0GTCCCCATGCTGGCCTCAGC 410550 5-10-5 389 22200 22219 0GGTCCCCATGCTGGCCTCAG 410551 5-10-5 390 22201 22220 0GGGTCCCCATGCTGGCCTCA 410552 5-10-5 391 22202 22221 0CGGGTCCCCATGCTGGCCTC 410553 5-10-5 392 22203 22222 0ACGGGTCCCCATGCTGGCCT 410554 5-10-5 393 22205 22224 0ACACGGGTCCCCATGCTGGC 410555 5-10-5 395 22207 22226 0GGACACGGGTCCCCATGCTG 410556 5-10-5 397 22209 22228 0GTGGACACGGGTCCCCATGC 410557 5-10-5 399 22211 22230 0CAGTGGACACGGGTCCCCAT 410558 5-10-5 401 22213 22232 0GGCAGTGGACACGGGTCCCC 410559 5-10-5 403 22215 22234 0GTGGCAGTGGACACGGGTCC 410560 5-10-5 404 22216 22235 0GGTGGCAGTGGACACGGGTC 410561 5-10-5 405 22217 22236 0TGGTGGCAGTGGACACGGGT 410562 5-10-5 411 24096 24115 0ACTTTGCATTCCAGACCTGG 410563 5-10-5 413 24098 24117 0TGACTTTGCATTCCAGACCT 410564 5-10-5 414 24099 24118 0TTGACTTTGCATTCCAGACC 410565 5-10-5 419 26112 26131 0CAGATGGCAACGGCTGTCAC 410566 5-10-5 420 26113 26132 0GCAGATGGCAACGGCTGTCA 410567 5-10-5 421 26114 26133 0AGCAGATGGCAACGGCTGTC 410568 5-10-5 422 26115 26134 0CAGCAGATGGCAACGGCTGT 410569 5-10-5 423 26116 26135 0GCAGCAGATGGCAACGGCTG 410570 5-10-5 424 26118 26137 0CGGCAGCAGATGGCAACGGC 410571 5-10-5 425 26119 26138 0CCGGCAGCAGATGGCAACGG 410572 5-10-5 427 26121 26140 0CTCCGGCAGCAGATGGCAAC 410573 5-10-5 428 26122 26141 0GCTCCGGCAGCAGATGGCAA 410730 5-10-5 202 6594 6613 0 CGCCACTCATCTTCACCAGG410731 5-10-5 207 6604 6623 0 TCCAGCAGGTCGCCACTCAT 410732 5-10-5 2149216 9235 0 GGGCTGGTATTCATCCGCCC 410733 5-10-5 231 14942 14961 0AAGTCGGTGACCATGACCCT 410734 5-10-5 279 18714 18733 0CCGGCAGCGGTGACCAGCAC 410735 5-10-5 291 19956 19975 0TCCCCAGGGTCACCGGCTGG 410736 5-10-5 303 19992 20011 0AGAGGTCCACACAGCGGCCA 410737 5-10-5 313 20042 20061 0GCAGGTGCTGCAGTCGCTGG 410738 5-10-5 324 20639 20658 0GCTCGGCAGACAGCATCATG 410739 5-10-5 336 20687 20706 0CAGAGAAGTGGATCAGTCTC 410740 5-10-5 345 20715 20734 0AACCAGGCCTCATTGATGAC 410741 5-10-5 369 21129 21148 0CCATCCGTGTAGGCCCCGAG 410742 5-10-5 159 2433 2452 0 GCCTGGAGCTGACGGTGCCC410743 5-10-5 170 2560 2579 0 TCCTCCTCGGAACGCAAGGC 410744 5-10-5 1712585 2604 0 GTGCTCGGGTGCTTCGGCCA 410745 5-10-5 172 2605 2624 0TGGAAGGTGGCTGTGGTTCC 410746 5-10-5 176 6444 6463 0 CGTAGGTGCCAGGCAACCTC410747 5-10-5 177 6482 6501 0 TGACTGCGAGAGGTGGGTCT 410748 5-10-5 1826528 6547 0 ATCCCCGGCGGGCAGCCTGG 410749 5-10-5 197 6565 6584 0AGAAGGCCATGGAAGACATG 410750 5-10-5 198 6575 6594 0 GAAGCCAGGAAGAAGGCCAT410752 5-10-5 209 9149 9168 0 GCAAAGACAGAGGAGTCCTC 410753 5-10-5 21514877 14896 0 ATACACCTCCACCAGGCTGC 410754 5-10-5 233 14954 14973 0GGCACATTCTCGAAGTCGGT 410756 5-10-5 235 15254 15273 0GGTGGGTGCCATGACTGTCA 410757 5-10-5 240 15279 15298 0TCCCGGCCGCTGACCACCCC 410758 5-10-5 256 15309 15328 0CGCATGCTGGCACCCTTGGC 410759 5-10-5 263 15358 15377 0GGTGCCGCTAACCGTGCCCT 410761 5-10-5 271 18614 18633 0GTGGCCCCACAGGCTGGACC 410762 5-10-5 272 18627 18646 0AGCAGCACCACCAGTGGCCC 410763 5-10-5 273 18649 18668 0GCTGTACCCACCCGCCAGGG 410764 5-10-5 274 18695 18714 0CGACCCCAGCCCTCGCCAGG 410767 5-10-5 289 19941 19960 0GCTGGTCTTGGGCATTGGTG 410768 5-10-5 305 20016 20035 0CAATGATGTCCTCCCCTGGG 410769 5-10-5 318 20061 20080 0TCCCACTCTGTGACACAAAG 410770 5-10-5 327 20657 20676 0CGGCCAGGGTGAGCTCCGGC 410771 5-10-5 360 20785 20804 0ACCTGCCCCATGGGTGCTGG 410772 5-10-5 364 21106 21125 0GCTGACCATACAGTCCTGCA 410773 5-10-5 387 22189 22208 0TGGCCTCAGCTGGTGGAGCT 410774 5-10-5 406 22220 22239 0TGTTGGTGGCAGTGGACACG 410776 5-10-5 408 23985 24004 0GTGCCAAGGTCCTCCACCTC 410777 5-10-5 409 24005 24024 0TCAGCACAGGCGGCTTGTGG 410778 5-10-5 410 24035 24054 0CCACGCACTGGTTGGGCTGA 410779 5-10-5 416 24115 24134 0CGGGATTCCATGCTCCTTGA 410781 5-10-5 418 25994 26013 0GGAGGGCACTGCAGCCAGTC 410782 5-10-5 429 26132 26151 0CCAGGTGCCGGCTCCGGCAG 410783 5-10-5 430 26142 26161 0GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 2. In certain such embodiments, thenucleotide sequences illustrated in Table 7 have a 3-14-3 gap-widenedmotif. Table 9 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 2, having a 3-14-3 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 9 Gapmer antisense compounds having a 3-14-3 motif targeted to SEQID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID IsisNo Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 399871 3-14-3 4 22742293 0 GCGCGGAATCCTGGCTGGGA 399872 3-14-3 5 2381 2400 0GAGGAGACCTAGAGGCCGTG 399873 3-14-3 6 2439 2458 0 AGGACCGCCTGGAGCTGACG399874 3-14-3 7 2549 2568 0 ACGCAAGGCTAGCACCAGCT 399875 3-14-3 9 26192638 0 CCTTGGCGCAGCGGTGGAAG 399876 3-14-3 10 6498 6517 0GGCGGGCAGTGCGCTCTGAC 399877 3-14-3 11 6537 6556 0 TGGTGAGGTATCCCCGGCGG399878 3-14-3 14 6557 6576 0 ATGGAAGACATGCAGGATCT 399879 3-14-3 15 65836602 0 TTCACCAGGAAGCCAGGAAG 399880 3-14-3 17 9130 9149 0CCTCGATGTAGTCGACATGG 399881 3-14-3 18 9210 9229 0 GTATTCATCCGCCCGGTACC399882 3-14-3 19 14891 14910 0 CTGGTGTCTAGGAGATACAC 399883 3-14-3 2114916 14935 0 GATTTCCCGGTGGTCACTCT 399884 3-14-3 24 14946 14965 0CTCGAAGTCGGTGACCATGA 399885 3-14-3 25 14979 14998 0 GTGGAAGCGGGTCCCGTCCT399886 3-14-3 26 15264 15283 0 ACCCCTGCCAGGTGGGTGCC 399887 3-14-3 2815298 15317 0 ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 15334 15353 0TTGGCAGTTGAGCACGCGCA 399890 3-14-3 32 18593 18612 0 GCTGGCTTTTCCGAATAAAC399891 3-14-3 33 18705 18724 0 GTGACCAGCACGACCCCAGC 399892 3-14-3 14919933 19952 0 TGGGCATTGGTGGCCCCAAC 399893 3-14-3 34 19962 19981 0CCAAAGTCCCCAGGGTCACC 399894 3-14-3 128 19966 19985 0GTCCCCAAAGTCCCCAGGGT 399895 3-14-3 36 19976 19995 0 GCCAAAGTTGGTCCCCAAAG399896 3-14-3 38 19997 20016 0 GGCAAAGAGGTCCACACAGC 399897 3-14-3 3920025 20044 0 TGGAGGCACCAATGATGTCC 399899 3-14-3 45 20691 20710 0TTGGCAGAGAAGTGGATCAG 399900 3-14-3 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC399901 3-14-3 87 20733 20752 0 ACCCGCTGGTCCTCAGGGAA 399902 3-14-3 11920762 20781 0 GCAGGGCGGCCACCAGGTTG 399904 3-14-3 54 21121 21140 0GTAGGCCCCGAGTGTGCTGA 399905 3-14-3 57 22096 22115 0 CGTTGTGGGCCCGGCAGACC399906 3-14-3 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT 399907 3-14-3 5922204 22223 0 CACGGGTCCCCATGCTGGCC 399908 3-14-3 60 24095 24114 0CTTTGCATTCCAGACCTGGG 399909 3-14-3 62 26117 26136 0 GGCAGCAGATGGCAACGGCT399910 3-14-3 154 26217 26236 0 TTTTAAAGCTCAGCCCCAGC 399911 3-14-3 6426311 26330 0 TCAAGGGCCAGGCCAGCAGC 399912 3-14-3 66 26404 26423 0ATGCCCCACAGTGAGGGAGG 399913 3-14-3 67 26413 26432 0 AATGGTGAAATGCCCCACAG399914 3-14-3 153 26456 26475 0 TTGGGAGCAGCTGGCAGCAC 399915 3-14-3 6826557 26576 0 CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 26639 26658 0GGAGATGAGGGCCATCAGCA 399917 3-14-3 70 26707 26726 0 TAGATGCCATCCAGAAAGCT399918 3-14-3 72 26790 26809 0 GGCATAGAGCAGAGTAAAGG 399919 3-14-3 11227034 27053 0 GAAGAGGCTTGGCTTCAGAG 399920 3-14-3 75 27244 27263 0GCTCAAGGAGGGACAGTTGT 399921 3-14-3 76 27279 27298 0 AAAGATAAATGTCTGCTTGC399922 3-14-3 78 27350 27369 0 TCTTCAAGTTACAAAAGCAA 399923 3-14-3 8513681 13700 0 ACAAATTCCCAGACTCAGCA 399924 3-14-3 129 13816 13835 0GTGCCATCTGAACAGCACCT 399925 3-14-3 110 13926 13945 0CCTGGAACCCCTGCAGCCAG 399926 3-14-3 152 13977 13996 0TTCAGGCAGGTTGCTGCTAG 399927 3-14-3 140 13998 14017 0TCAGCCAGGCCAAAGGAAGA 399928 3-14-3 136 14122 14141 0TAGGAGAAAGTAGGGAGAGC 399929 3-14-3 132 14179 14198 0TAAAAGCTGCAAGAGACTCA 399930 3-14-3 139 14267 14286 0TCAGAGAAAACAGTCACCGA 399931 3-14-3 142 14404 14423 0TCATTTTAGAGACAGGAAGC 399932 3-14-3 113 14441 14460 0GAATAACAGTGATGTCTGGC 399933 3-14-3 138 14494 14513 0TCACAGCTCACCGAGTCTGC 399934 3-14-3 98 14524 14543 0 AGTGTAAAATAAAGCCCCTA399935 3-14-3 96 14601 14620 0 AGGACCCAAGTCATCCTGCT 399936 3-14-3 12414631 14650 0 GGCCATCAGCTGGCAATGCT 399937 3-14-3 133 14675 14694 0TAGACAAGGAAAGGGAGGCC 399938 3-14-3 103 14681 14700 0ATTTCATAGACAAGGAAAGG 399939 3-14-3 155 14801 14820 0CTTATAGTTAACACACAGAA 399943 3-14-3 102 5590 5609 0 ATGTGCAGAGATCAATCACA399944 3-14-3 127 10633 10652 0 GGTGGTAATTTGTCACAGCA 399945 3-14-3 8411308 11327 0 AAGGTCACACAGTTAAGAGT 399946 3-14-3 157 18561 18580 0AGGAACAAAGCCAAGGTCAC 399947 3-14-3 126 22292 22311 0GGTGCATAAGGAGAAAGAGA 399948 3-14-3 147 24858 24877 0TGAGTTCATTTAAGAGTGGA 399949 3-14-3 8 2556 2575 0 CCTCGGAACGCAAGGCTAGC399950 3-14-3 12 6543 6562 0 GGATCTTGGTGAGGTATCCC 399951 3-14-3 13 65526571 0 AGACATGCAGGATCTTGGTG 399952 3-14-3 16 6588 6607 0TCATCTTCACCAGGAAGCCA 399953 3-14-3 20 14896 14915 0 GTATGCTGGTGTCTAGGAGA399954 3-14-3 22 14922 14941 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 2314936 14955 0 GTGACCATGACCCTGCCCTC 399956 3-14-3 27 15269 15288 0TGACCACCCCTGCCAGGTGG 399957 3-14-3 30 15339 15358 0 TTCCCTTGGCAGTTGAGCAC399958 3-14-3 35 19971 19990 0 AGTTGGTCCCCAAAGTCCCC 399959 3-14-3 3719988 20007 0 GTCCACACAGCGGCCAAAGT 399960 3-14-3 40 20036 20055 0GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 20047 20066 0 ACAAAGCAGGTGCTGCAGTC399963 3-14-3 43 20635 20654 0 GGCAGACAGCATCATGGCTG 399964 3-14-3 4420683 20702 0 GAAGTGGATCAGTCTCTGCC 399965 3-14-3 46 20696 20715 0CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 20702 20721 0 TGATGACATCTTTGGCAGAG399967 3-14-3 48 20709 20728 0 GCCTCATTGATGACATCTTT 399968 3-14-3 4920721 20740 0 TCAGGGAACCAGGCCTCATT 399969 3-14-3 51 20740 20759 0GGTCAGTACCCGCTGGTCCT 399970 3-14-3 53 21091 21110 0 CTGCAAAACAGCTGCCAACC399971 3-14-3 55 21186 21205 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 5621192 21211 0 TCCTGGAGAAACTGGAGCAG 399973 3-14-3 61 24100 24119 0CTTGACTTTGCATTCCAGAC 399974 3-14-3 63 26222 26241 0 AACCATTTTAAAGCTCAGCC399975 3-14-3 65 26316 26335 0 CCCACTCAAGGGCCAGGCCA 399976 3-14-3 12226389 26408 0 GGAGGGAGCTTCCTGGCACC 399977 3-14-3 71 26713 26732 0TCTGGCTAGATGCCATCCAG 399978 3-14-3 73 26795 26814 0 AGCCTGGCATAGAGCAGAGT399979 3-14-3 135 26801 26820 0 TAGCACAGCCTGGCATAGAG 399980 3-14-3 7427040 27059 0 AAGTAAGAAGAGGCTTGGCT 399981 3-14-3 77 27284 27303 0ACCCAAAAGATAAATGTCTG 399982 3-14-3 99 27357 27376 0 ATAAATATCTTCAAGTTACA399983 3-14-3 100 13746 13765 0 ATCTCAGGACAGGTGAGCAA 399984 3-14-3 11613760 13779 0 GAGTAGAGATTCTCATCTCA 399985 3-14-3 117 13828 13847 0GAGTCTTCTGAAGTGCCATC 399986 3-14-3 81 13903 13922 0 AAGCAGGGCCTCAGGTGGAA399987 3-14-3 83 13986 14005 0 AAGGAAGACTTCAGGCAGGT 399988 3-14-3 13714112 14131 0 TAGGGAGAGCTCACAGATGC 399989 3-14-3 92 14397 14416 0AGAGACAGGAAGCTGCAGCT 399990 3-14-3 82 14670 14689 0 AAGGAAAGGGAGGCCTAGAG399991 3-14-3 156 14809 14828 0 AAGTCAACCTTATAGTTAAC 399993 3-14-3 1073056 3075 0 CCCACTATAATGGCAAGCCC 399994 3-14-3 80 4306 4325 0AACCCAGTTCTAATGCACCT 399995 3-14-3 106 5140 5159 0 CCAGTCAGAGTAGAACAGAG399996 3-14-3 121 5599 5618 0 GGAGCCTACATGTGCAGAGA 399997 3-14-3 94 56675686 0 AGCATGGCACCAGCATCTGC 399998 3-14-3 108 6652 6671 0CCCAGCCCTATCAGGAAGTG 399999 3-14-3 144 7099 7118 0 TGACATCCAGGAGGGAGGAG400000 3-14-3 91 7556 7575 0 AGACTGATGGAAGGCATTGA 400001 3-14-3 131 75657584 0 GTGTTGAGCAGACTGATGGA 400002 3-14-3 145 8836 8855 0TGACATCTTGTCTGGGAGCC 400003 3-14-3 90 8948 8967 0 AGACTAGGAGCCTGAGTTTT400004 3-14-3 125 9099 9118 0 GGCCTGCAGAAGCCAGAGAG 400005 3-14-3 14810252 10271 0 TGGCAGCAACTCAGACATAT 400006 3-14-3 79 11472 11491 0AAATGCAGGGCTAAAATCAC 400007 3-14-3 88 12715 12734 0 ACTGGATACATTGGCAGACA400008 3-14-3 111 12928 12947 0 CTAGAGGAACCACTAGATAT 400009 3-14-3 8615471 15490 0 ACAGCATTCTTGGTTAGGAG 400010 3-14-3 97 16134 16153 0AGTCAAGCTGCTGCCCAGAG 400011 3-14-3 120 16668 16687 0GCTAGTTATTAAGCACCTGC 400012 3-14-3 150 17267 17286 0TGTGAGCTCTGGCCCAGTGG 400013 3-14-3 115 18377 18396 0GAGTAAGGCAGGTTACTCTC 400014 3-14-3 134 18408 18427 0TAGATGTGACTAACATTTAA 400015 3-14-3 105 19203 19222 0CACATTAGCCTTGCTCAAGT 400016 3-14-3 151 19913 19932 0TGTGATGACCTGGAAAGGTG 400017 3-14-3 158 20100 20119 0GTGGTGACTTACCAGCCACG 400018 3-14-3 109 20188 20207 0CCCCTGCACAGAGCCTGGCA 400019 3-14-3 141 20624 20643 0TCATGGCTGCAATGCCTGGT 400020 3-14-3 93 20995 21014 0 AGAGAGGAGGGCTTAAAGAA400021 3-14-3 95 21082 21101 0 AGCTGCCAACCTGCAAAAAG 400022 3-14-3 14321481 21500 0 TGAAAATCCATCCAGCACTG 400023 3-14-3 89 21589 21608 0AGAACCATGGAGCACCTGAG 400024 3-14-3 123 21696 21715 0GGCACTGCCCTTCCACCAAA 400025 3-14-3 118 24907 24926 0GCACCATCCAGACCAGAATC 400026 3-14-3 114 25413 25432 0GAGAGGTTCAGATCCAGGCC 405604 3-14-3 236 15257 15276 0CCAGGTGGGTGCCATGACTG 405641 3-14-3 373 21183 21202 0AACTGGAGCAGCTCAGCAGC 410574 3-14-3 184 6534 6553 0 TGAGGTATCCCCGGCGGGCA410575 3-14-3 185 6535 6554 0 GTGAGGTATCCCCGGCGGGC 410576 3-14-3 1866536 6555 0 GGTGAGGTATCCCCGGCGGG 410577 3-14-3 187 6538 6557 0TTGGTGAGGTATCCCCGGCG 410578 3-14-3 188 6539 6558 0 CTTGGTGAGGTATCCCCGGC410579 3-14-3 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG 410580 3-14-3 1906541 6560 0 ATCTTGGTGAGGTATCCCCG 410581 3-14-3 191 6542 6561 0GATCTTGGTGAGGTATCCCC 410582 3-14-3 192 6544 6563 0 AGGATCTTGGTGAGGTATCC410583 3-14-3 243 15291 15310 0 GCCACGCCGGCATCCCGGCC 410584 3-14-3 24415292 15311 0 GGCCACGCCGGCATCCCGGC 410585 3-14-3 245 15293 15312 0TGGCCACGCCGGCATCCCGG 410586 3-14-3 246 15294 15313 0TTGGCCACGCCGGCATCCCG 410587 3-14-3 247 15295 15314 0CTTGGCCACGCCGGCATCCC 410588 3-14-3 248 15296 15315 0CCTTGGCCACGCCGGCATCC 410589 3-14-3 249 15297 15316 0CCCTTGGCCACGCCGGCATC 410590 3-14-3 250 15299 15318 0CACCCTTGGCCACGCCGGCA 410591 3-14-3 251 15300 15319 0GCACCCTTGGCCACGCCGGC 410592 3-14-3 252 15301 15320 0GGCACCCTTGGCCACGCCGG 410593 3-14-3 253 15302 15321 0TGGCACCCTTGGCCACGCCG 410594 3-14-3 254 15303 15322 0CTGGCACCCTTGGCCACGCC 410595 3-14-3 255 15304 15323 0GCTGGCACCCTTGGCCACGC 410596 3-14-3 348 20719 20738 0AGGGAACCAGGCCTCATTGA 410597 3-14-3 349 20720 20739 0CAGGGAACCAGGCCTCATTG 410598 3-14-3 350 20722 20741 0CTCAGGGAACCAGGCCTCAT 410599 3-14-3 351 20723 20742 0CCTCAGGGAACCAGGCCTCA 410600 3-14-3 352 20724 20743 0TCCTCAGGGAACCAGGCCTC 410601 3-14-3 353 20725 20744 0GTCCTCAGGGAACCAGGCCT 410602 3-14-3 354 20727 20746 0TGGTCCTCAGGGAACCAGGC 410603 3-14-3 355 20728 20747 0CTGGTCCTCAGGGAACCAGG 410604 3-14-3 356 20729 20748 0GCTGGTCCTCAGGGAACCAG 410605 3-14-3 376 22133 22152 0CACCTGGCAATGGCGTAGAC 410606 3-14-3 377 22134 22153 0GCACCTGGCAATGGCGTAGA 410607 3-14-3 378 22135 22154 0AGCACCTGGCAATGGCGTAG 410608 3-14-3 379 22136 22155 0CAGCACCTGGCAATGGCGTA 410609 3-14-3 380 22137 22156 0GCAGCACCTGGCAATGGCGT 410610 3-14-3 381 22138 22157 0GGCAGCACCTGGCAATGGCG 410611 3-14-3 382 22139 22158 0AGGCAGCACCTGGCAATGGC 410612 3-14-3 383 22140 22159 0CAGGCAGCACCTGGCAATGG 410613 3-14-3 384 22141 22160 0GCAGGCAGCACCTGGCAATG 410614 3-14-3 385 22143 22162 0TAGCAGGCAGCACCTGGCAA 410615 3-14-3 388 22199 22218 0GTCCCCATGCTGGCCTCAGC 410616 3-14-3 389 22200 22219 0GGTCCCCATGCTGGCCTCAG 410617 3-14-3 390 22201 22220 0GGGTCCCCATGCTGGCCTCA 410618 3-14-3 391 22202 22221 0CGGGTCCCCATGCTGGCCTC 410619 3-14-3 392 22203 22222 0ACGGGTCCCCATGCTGGCCT 410620 3-14-3 393 22205 22224 0ACACGGGTCCCCATGCTGGC 410621 3-14-3 394 22206 22225 0GACACGGGTCCCCATGCTGG 410622 3-14-3 395 22207 22226 0GGACACGGGTCCCCATGCTG 410623 3-14-3 396 22208 22227 0TGGACACGGGTCCCCATGCT 410624 3-14-3 397 22209 22228 0GTGGACACGGGTCCCCATGC 410625 3-14-3 398 22210 22229 0AGTGGACACGGGTCCCCATG 410626 3-14-3 399 22211 22230 0CAGTGGACACGGGTCCCCAT 410627 3-14-3 400 22212 22231 0GCAGTGGACACGGGTCCCCA 410628 3-14-3 401 22213 22232 0GGCAGTGGACACGGGTCCCC 410629 3-14-3 402 22214 22233 0TGGCAGTGGACACGGGTCCC 410630 3-14-3 403 22215 22234 0GTGGCAGTGGACACGGGTCC 410631 3-14-3 404 22216 22235 0GGTGGCAGTGGACACGGGTC 410632 3-14-3 405 22217 22236 0TGGTGGCAGTGGACACGGGT 410633 3-14-3 411 24096 24115 0ACTTTGCATTCCAGACCTGG 410634 3-14-3 412 24097 24116 0GACTTTGCATTCCAGACCTG 410635 3-14-3 413 24098 24117 0TGACTTTGCATTCCAGACCT 410636 3-14-3 414 24099 24118 0TTGACTTTGCATTCCAGACC 410637 3-14-3 419 26112 26131 0CAGATGGCAACGGCTGTCAC 410638 3-14-3 420 26113 26132 0GCAGATGGCAACGGCTGTCA 410639 3-14-3 421 26114 26133 0AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 26115 26134 0CAGCAGATGGCAACGGCTGT 410641 3-14-3 423 26116 26135 0GCAGCAGATGGCAACGGCTG 410642 3-14-3 424 26118 26137 0CGGCAGCAGATGGCAACGGC 410643 3-14-3 425 26119 26138 0CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 26120 26139 0TCCGGCAGCAGATGGCAACG 410645 3-14-3 427 26121 26140 0CTCCGGCAGCAGATGGCAAC 410646 3-14-3 428 26122 26141 0GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 2. In certain such embodiments, thenucleotide sequences illustrated in Table 7 have a 2-13-5 gap-widenedmotif. Table 10 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 2, having a 2-13-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 10 Gapmer antisense compounds having a 2-13-5 motif targeted toSEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ IDIsis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 410647 2-13-5184 6534 6553 0 TGAGGTATCCCCGGCGGGCA 410648 2-13-5 185 6535 6554 0GTGAGGTATCCCCGGCGGGC 410649 2-13-5 186 6536 6555 0 GGTGAGGTATCCCCGGCGGG410650 2-13-5 11 6537 6556 0 TGGTGAGGTATCCCCGGCGG 410651 2-13-5 187 65386557 0 TTGGTGAGGTATCCCCGGCG 410652 2-13-5 188 6539 6558 0CTTGGTGAGGTATCCCCGGC 410653 2-13-5 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG410654 2-13-5 190 6541 6560 0 ATCTTGGTGAGGTATCCCCG 410655 2-13-5 1916542 6561 0 GATCTTGGTGAGGTATCCCC 410656 2-13-5 12 6543 6562 0GGATCTTGGTGAGGTATCCC 410657 2-13-5 192 6544 6563 0 AGGATCTTGGTGAGGTATCC410658 2-13-5 243 15291 15310 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 24415292 15311 0 GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 15293 15312 0TGGCCACGCCGGCATCCCGG 410661 2-13-5 246 15294 15313 0TTGGCCACGCCGGCATCCCG 410662 2-13-5 247 15295 15314 0CTTGGCCACGCCGGCATCCC 410663 2-13-5 248 15296 15315 0CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 15297 15316 0CCCTTGGCCACGCCGGCATC 410665 2-13-5 28 15298 15317 0 ACCCTTGGCCACGCCGGCAT410666 2-13-5 250 15299 15318 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 25115300 15319 0 GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 15301 15320 0GGCACCCTTGGCCACGCCGG 410669 2-13-5 253 15302 15321 0TGGCACCCTTGGCCACGCCG 410670 2-13-5 254 15303 15322 0CTGGCACCCTTGGCCACGCC 410671 2-13-5 255 15304 15323 0GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 20719 20738 0AGGGAACCAGGCCTCATTGA 410673 2-13-5 349 20720 20739 0CAGGGAACCAGGCCTCATTG 410674 2-13-5 49 20721 20740 0 TCAGGGAACCAGGCCTCATT410675 2-13-5 350 20722 20741 0 CTCAGGGAACCAGGCCTCAT 410676 2-13-5 35120723 20742 0 CCTCAGGGAACCAGGCCTCA 410677 2-13-5 352 20724 20743 0TCCTCAGGGAACCAGGCCTC 410678 2-13-5 353 20725 20744 0GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC410680 2-13-5 354 20727 20746 0 TGGTCCTCAGGGAACCAGGC 410681 2-13-5 35520728 20747 0 CTGGTCCTCAGGGAACCAGG 410682 2-13-5 356 20729 20748 0GCTGGTCCTCAGGGAACCAG 410683 2-13-5 376 22133 22152 0CACCTGGCAATGGCGTAGAC 410684 2-13-5 377 22134 22153 0GCACCTGGCAATGGCGTAGA 410685 2-13-5 378 22135 22154 0AGCACCTGGCAATGGCGTAG 410686 2-13-5 379 22136 22155 0CAGCACCTGGCAATGGCGTA 410687 2-13-5 380 22137 22156 0GCAGCACCTGGCAATGGCGT 410688 2-13-5 381 22138 22157 0GGCAGCACCTGGCAATGGCG 410689 2-13-5 382 22139 22158 0AGGCAGCACCTGGCAATGGC 410690 2-13-5 383 22140 22159 0CAGGCAGCACCTGGCAATGG 410691 2-13-5 384 22141 22160 0GCAGGCAGCACCTGGCAATG 410692 2-13-5 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT410693 2-13-5 385 22143 22162 0 TAGCAGGCAGCACCTGGCAA 410694 2-13-5 38822199 22218 0 GTCCCCATGCTGGCCTCAGC 410695 2-13-5 389 22200 22219 0GGTCCCCATGCTGGCCTCAG 410696 2-13-5 390 22201 22220 0GGGTCCCCATGCTGGCCTCA 410697 2-13-5 391 22202 22221 0CGGGTCCCCATGCTGGCCTC 410698 2-13-5 392 22203 22222 0ACGGGTCCCCATGCTGGCCT 410699 2-13-5 59 22204 22223 0 CACGGGTCCCCATGCTGGCC410700 2-13-5 393 22205 22224 0 ACACGGGTCCCCATGCTGGC 410701 2-13-5 39422206 22225 0 GACACGGGTCCCCATGCTGG 410702 2-13-5 395 22207 22226 0GGACACGGGTCCCCATGCTG 410703 2-13-5 396 22208 22227 0TGGACACGGGTCCCCATGCT 410704 2-13-5 397 22209 22228 0GTGGACACGGGTCCCCATGC 410705 2-13-5 398 22210 22229 0AGTGGACACGGGTCCCCATG 410706 2-13-5 399 22211 22230 0CAGTGGACACGGGTCCCCAT 410707 2-13-5 400 22212 22231 0GCAGTGGACACGGGTCCCCA 410708 2-13-5 401 22213 22232 0GGCAGTGGACACGGGTCCCC 410709 2-13-5 402 22214 22233 0TGGCAGTGGACACGGGTCCC 410710 2-13-5 403 22215 22234 0GTGGCAGTGGACACGGGTCC 410711 2-13-5 404 22216 22235 0GGTGGCAGTGGACACGGGTC 410712 2-13-5 405 22217 22236 0TGGTGGCAGTGGACACGGGT 410713 2-13-5 60 24095 24114 0 CTTTGCATTCCAGACCTGGG410714 2-13-5 411 24096 24115 0 ACTTTGCATTCCAGACCTGG 410715 2-13-5 41224097 24116 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 413 24098 24117 0TGACTTTGCATTCCAGACCT 410717 2-13-5 414 24099 24118 0TTGACTTTGCATTCCAGACC 410718 2-13-5 61 24100 24119 0 CTTGACTTTGCATTCCAGAC410719 2-13-5 419 26112 26131 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 42026113 26132 0 GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 26114 26133 0AGCAGATGGCAACGGCTGTC 410722 2-13-5 422 26115 26134 0CAGCAGATGGCAACGGCTGT 410723 2-13-5 423 26116 26135 0GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 26117 26136 0 GGCAGCAGATGGCAACGGCT410725 2-13-5 424 26118 26137 0 CGGCAGCAGATGGCAACGGC 410726 2-13-5 42526119 26138 0 CCGGCAGCAGATGGCAACGG 410727 2-13-5 426 26120 26139 0TCCGGCAGCAGATGGCAACG 410728 2-13-5 427 26121 26140 0CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 26122 26141 0GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 2. In certain such embodiments, thenucleotide sequences illustrated in Table 7 have a 3-13-4 gap-widenedmotif. Table 11 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 2, having a 3-13-4 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 11 Gapmer antisense compounds having a 3-13-4 motif targeted toSEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ IDIsis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 405526 3-13-4236 15257 15276 0 CCAGGTGGGTGCCATGACTG 405557 3-13-4 50 20726 20745 0GGTCCTCAGGGAACCAGGCC 405564 3-13-4 373 21183 21202 0AACTGGAGCAGCTCAGCAGC

The following embodiments set forth target regions of PCSK9 nucleicacids. Also illustrated are examples of antisense compounds targeted tothe target regions. It is understood that the sequence set forth in eachSEQ ID NO is independent of any modification to a sugar moiety, aninternucleoside linkage, or a nucleobase. As such, antisense compoundsdefined by a SEQ ID NO may comprise, independently, one or moremodifications to a sugar moiety, an internucleoside linkage, or anucleobase. Antisense compounds described by Isis Number (Isis No)indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575,2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638,3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463,6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562,6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567,6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621,6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967,9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235,10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947,13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141,14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513,14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700,14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943,14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998,15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318,15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322,15296-15315, 15296-15323, 15297-15323, 15298-15323, 15299-15323,15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366,15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427,18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746,18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952,19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016,20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119,20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660,20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706,20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744,20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749,20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107,21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209,21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719,22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160,22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163,22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227,22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232,22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236,22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054,24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432,25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140,26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475,26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241,26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734,26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.

In certain embodiments, a target region is nucleotides 2274-2400 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2274-2400 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 4 or 5. In certain suchembodiments, an antisense compound targeted to nucleotides 2274-2400 ofSEQ ID NO: 2 is selected from ISIS NOs: 395149, 399871, 395150 or399872.

In certain embodiments, a target region is nucleotides 2274-2575 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2274-2575 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 4, 5, 6, 7, 8, 159, 160,162, 163, 164, 165, 166, 167, 168 or 169. In certain such embodiments,an antisense compound targeted to nucleotides 2274-2575 of SEQ ID NO: 2is selected from ISIS NOs: 395149, 399871, 395150, 399872, 410742,405999, 395151, 399873, 405861, 405862, 405863, 405864, 395152, 399874,405865, 405866, 405867, 405868, 399793, or 399949.

In certain embodiments, a target region is nucleotides 2433-2570 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2433-2570 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 6, 7, 159, 160, 162, 163,164, 165, 166, or 167. In certain such embodiments, an antisensecompound targeted to nucleotides 2433-2570 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395151, 395152, 399873, 399874, 405861, 405862, 405863,405864, 405865, 405866, 405999, or 410742.

In certain embodiments, a target region is nucleotides 2433-2579 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2433-2579 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 6, 7, 8, 159, 160, 162,163, 164, 165, 166, 167, 168, 169, or 170. In certain such embodiments,an antisense compound targeted to nucleotides 2433-2579 of SEQ ID NO: 2is selected from ISIS NOs: 395151, 395152, 399793, 399873, 399874,399949, 405861, 405862, 405863, 405864, 405865, 405866, 405867, 405868,405999, 410742, or 410743.

In certain embodiments, a target region is nucleotides 2549-2575 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2549-2575 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168 or169. In certain such embodiments, an antisense compound targeted tonucleotides 2549-2575 of SEQ ID NO: 2 is selected from ISIS NOs: 395152,399793, 399874, 399949, 405865, 405866, 405867, or 405868.

In certain embodiments, a target region is nucleotides 2552-2579 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2552-2579 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 8, 168, 169 or 170. Incertain such embodiments, an antisense compound targeted to nucleotides2552-2579 of SEQ ID NO: 2 is selected from ISIS NOs: 399793, 399949,405867, 405868, or 410743.

In certain embodiments, a target region is nucleotides 2585-2638 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2585-2638 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 9, 171 or 172. In certainsuch embodiments, an antisense compound targeted to nucleotides2585-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 395153, 399875,410744, or 410745.

In certain embodiments, a target region is nucleotides 2605-2638 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 2605-2638 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 8, 168 or 169. In certainsuch embodiments, an antisense compound targeted to nucleotides2605-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 410745, 395153, or399875.

In certain embodiments, a target region is nucleotides 3056-3075 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 3056-3075 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 107. In certain suchembodiments, an antisense compound targeted to nucleotides 3056-3075 ofSEQ ID NO: 2 is selected from ISIS NOs: 399837 or 399993.

In certain embodiments, a target region is nucleotides 4150-5159 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 4150-5159 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 106. In certain suchembodiments, an antisense compound targeted to nucleotides 4150-5159 ofSEQ ID NO: 2 is selected from ISIS NOs: 399839 or 399995.

In certain embodiments, a target region is nucleotides 4306-4325 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 4306-4325 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 80. In certain suchembodiments, an antisense compound targeted to nucleotides 4306-4325 ofSEQ ID NO: 2 is selected from ISIS NOs: 399838 or 399994.

In certain embodiments, a target region is nucleotides 5590-5618 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 5590-5618 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 102 or 121. In certainsuch embodiments, an antisense compound targeted to nucleotides5590-5618 of SEQ ID NO: 2 is selected from ISIS NOs: 395221, 399840,399943, or 399996.

In certain embodiments, a target region is nucleotides 5667-5686 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 5667-5686 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 94. In certain suchembodiments, an antisense compound targeted to nucleotides 5667-5686 ofSEQ ID NO: 2 is selected from ISIS NOs: 399841 or 399997.

In certain embodiments, a target region is nucleotides 6444-6463 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6444-6463 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 176. In certain suchembodiments, an antisense compound targeted to nucleotides 6444-6463 ofSEQ ID NO: 2 is selected from ISIS NOs: 410746.

In certain embodiments, a target region is nucleotides 6482-6518 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6482-6518 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or181. In certain such embodiments, an antisense compound targeted tonucleotides 6482-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154,399876, 406003, 406004, 406005, 406006 or 410747.

In certain embodiments, a target region is nucleotides 6492-6518 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6492-6518 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 10, 178, 179, 180 or 181.In certain such embodiments, an antisense compound targeted tonucleotides 6492-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154,399876, 406003, 406004, 406005 or 406006.

In certain embodiments, a target region is nucleotides 6528-6555 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6528-6555 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 182, 183, 184, 185 or 186.In certain such embodiments, an antisense compound targeted tonucleotides 6528-6555 of SEQ ID NO: 2 is selected from ISIS NOs: 406007,410529, 410530, 410531, 410574, 410575, 410576, 410647, 410648, 410649or 410748.

In certain embodiments, a target region is nucleotides 6528-6623 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6528-6623 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 13, 14, 15,16, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, or 207.In certain such embodiments, an antisense compound targeted tonucleotides 6528-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 395155,395156, 395157, 399794, 399795, 399796, 399877, 399878, 399879, 399950,399951, 399952, 406007, 406008, 406009, 406010, 406011, 406012, 406013,406014, 406015, 406016, 406017, 406018, 406019, 406020, 410529, 410530,410531, 410532, 410533, 410534, 410535, 410574, 410575, 410576, 410577,410578, 410579, 410580, 410581, 410582, 410647, 410648, 410649, 410650,410651, 410652, 410653, 410654, 410655, 410656, 410657, 410730, 410731,410748, 410749, or 410750.

In certain embodiments, a target region is nucleotides 6534-6561 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6534-6561 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 184, 185, 186, 187,188, 189, 190 or 191. In certain such embodiments, an antisense compoundtargeted to nucleotides 6534-6561 of SEQ ID NO: 2 is selected from ISISNOs: 395155, 399877, 406008, 406009, 410529, 410530, 410531, 410532,410533, 410534, 410574, 410575, 410576, 410577, 410578, 410579, 410580,410581, 410647, 410648, 410649, 410650, 410651, 410652, 410653, 410654,or 410655.

In certain embodiments, a target region is nucleotides 6535-6562 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6535-6562 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 185, 186, 187,188, 189, 190 or 191. In certain such embodiments, an antisense compoundtargeted to nucleotides 6535-6562 of SEQ ID NO: 2 is selected from ISISNOs: 395155, 399794, 399877, 399950, 406008, 406009, 410530, 410531,410532, 410533, 410534, 410575, 410576, 410577, 410578, 410579, 410580,410581, 410648, 410649, 410650, 410651, 410652, 410653, 410654, 410655or 410656.

In certain embodiments, a target region is nucleotides 6536-6563 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6536-6563 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 186, 187, 188,189, 190, 191 or 192. In certain such embodiments, an antisense compoundtargeted to nucleotides 6536-6563 of SEQ ID NO: 2 is selected from ISISNOs: 395155, 399794, 399877, 399950, 406008, 406009, 410531, 410532,410533, 410534, 410535, 410576, 410577, 410578, 410579, 410580, 410581,410582, 410649, 410650, 410651, 410652, 410653, 410654, 410655, 410656or 410657.

In certain embodiments, a target region is nucleotides 6537-6563 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6537-6563 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 11, 12, 187, 188, 189,190, 191 or 192. In certain such embodiments, an antisense compoundtargeted to nucleotides 6537-6563 of SEQ ID NO: 2 is selected from ISISNOs: 395155, 399794, 399877, 399950, 406008, 406009, 410532, 410533,410534, 410535, 410577, 410578, 410579, 410580, 410581, 410582, 410650,410651, 410652, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6538-6565 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6538-6565 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190,191, 192 or 193. In certain such embodiments, an antisense compoundtargeted to nucleotides 6538-6565 of SEQ ID NO: 2 is selected from ISISNOs: 399794, 399950, 406008, 406009, 406010, 410532, 410533, 410534,410535, 410577, 410578, 410579, 410580, 410581, 410582, 410651, 410652,410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6539-6565 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6539-6565 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191,192 or 193. In certain such embodiments, an antisense compound targetedto nucleotides 6539-6565 of SEQ ID NO: 2 is selected from ISIS NOs:399794, 399950, 406008, 406009, 406010, 410533, 410534, 410535, 410578,410579, 410580, 410581, 410582, 410652, 410653, 410654, 410655, 410656or 410657.

In certain embodiments, a target region is nucleotides 6540-6567 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6540-6567 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192,193 or 194. In certain such embodiments, an antisense compound targetedto nucleotides 6540-6567 of SEQ ID NO: 2 is selected from ISIS NOs:399794, 399950, 406009, 406010, 406011, 410533, 410534, 410535, 410579,410580, 410581, 410582, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6541-6567 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6541-6567 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 190, 191, 192, 193 or194. In certain such embodiments, an antisense compound targeted tonucleotides 6541-6567 of SEQ ID NO: 2 is selected from ISIS NOs: 399794,399950, 406009, 406010, 406011, 410534, 410535, 410580, 410581, 410582,410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6542-6569 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6542-6569 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 12, 191, 192, 193, 194 or195. In certain such embodiments, an antisense compound targeted tonucleotides 6542-6569 of SEQ ID NO: 2 is selected from ISIS NOs: 399794,399950, 406010, 406011, 406012, 410534, 410535, 410581, 410582, 410655,410656 or 410657.

In certain embodiments, a target region is nucleotides 6546-6573 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6546-6573 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 13, 193, 194, 195 or 196.In certain such embodiments, an antisense compound targeted tonucleotides 6546-6573 of SEQ ID NO: 2 is selected from ISIS NOs: 399795,399951, 406010, 406011, 406012 or 406013.

In certain embodiments, a target region is nucleotides 6557-6584 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6557-6584 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 14, 197 or 198. In certainsuch embodiments, an antisense compound targeted to nucleotides6557-6584 of SEQ ID NO: 2 is selected from ISIS NOs: 395156, 399878,410749 or 410750.

In certain embodiments, a target region is nucleotides 6575-6602 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6575-6602 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain suchembodiments, an antisense compound targeted to nucleotides 6575-6602 ofSEQ ID NO: 2 is selected from ISIS NOs: 395157, 399879 or 410750.

In certain embodiments, a target region is nucleotides 6585-6611 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6585-6611 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 16, 199, 200 or 201. Incertain such embodiments, an antisense compound targeted to nucleotides6585-6611 of SEQ ID NO: 2 is selected from ISIS NOs: 399796, 399952,406014, 406015 or 406016.

In certain embodiments, a target region is nucleotides 6594-6621 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6594-6621 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205 or 206.In certain such embodiments, an antisense compound targeted tonucleotides 6594-6621 of SEQ ID NO: 2 is selected from ISIS NOs: 406017,406018, 406019, 406020 or 410730.

In certain embodiments, a target region is nucleotides 6596-6623 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6596-6623 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 203, 204, 205, 206 or 207.In certain such embodiments, an antisense compound targeted tonucleotides 6596-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 406017,406018, 406019, 406020 or 410731.

In certain embodiments, a target region is nucleotides 6652-6671 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 6652-6671 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 108. In certain suchembodiments, an antisense compound targeted to nucleotides 6652-6671 ofSEQ ID NO: 2 is selected from ISIS NOs: 399842 or 399998.

In certain embodiments, a target region is nucleotides 7099-7118 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 7099-7118 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 114. In certain suchembodiments, an antisense compound targeted to nucleotides 7099-7118 ofSEQ ID NO: 2 is selected from ISIS NOs: 399843 or 399999.

In certain embodiments, a target region is nucleotides 7556-7584 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 7556-7584 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 91 or 131. In certain suchembodiments, an antisense compound targeted to nucleotides 7556-7584 ofSEQ ID NO: 2 is selected from ISIS NOs: 399844, 399845, 400000 or400001.

In certain embodiments, a target region is nucleotides 8836-8855 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 8836-8855 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 145. In certain suchembodiments, an antisense compound targeted to nucleotides 8836-8855 ofSEQ ID NO: 2 is selected from ISIS NOs: 399846 or 400002.

In certain embodiments, a target region is nucleotides 8948-8967 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 8948-8967 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 90. In certain suchembodiments, an antisense compound targeted to nucleotides 8948-8967 ofSEQ ID NO: 2 is selected from ISIS NOs: 399847 or 400003.

In certain embodiments, a target region is nucleotides 9099-9118 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9099-9118 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 125. In certain suchembodiments, an antisense compound targeted to nucleotides 9099-9118 ofSEQ ID NO: 2 is selected from ISIS NOs: 399848 or 400004.

In certain embodiments, a target region is nucleotides 9099-9168 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9099-9168 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 17, 125 or 209. In certainsuch embodiments, an antisense compound targeted to nucleotides9099-9168 of SEQ ID NO: 2 is selected from ISIS NOs: 395158, 399848,399880, 400004 or 410752.

In certain embodiments, a target region is nucleotides 9130-9168 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9130-9168 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 17. In certain suchembodiments, an antisense compound targeted to nucleotides 9130-9168 ofSEQ ID NO: 2 is selected from ISIS NOs: 395158 or 399880.

In certain embodiments, a target region is nucleotides 9207-9233 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9207-9233 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212 or 213.In certain such embodiments, an antisense compound targeted tonucleotides 9207-9233 of SEQ ID NO: 2 is selected from ISIS NOs: 395159,399881, 406021, 406022, 406023 or 406024.

In certain embodiments, a target region is nucleotides 9207-9235 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9207-9235 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213 or214. In certain such embodiments, an antisense compound targeted tonucleotides 9207-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159,399881, 406021, 406022, 406023, 406024 or 410732.

In certain embodiments, a target region is nucleotides 9209-9235 of SEQID NO: 2. In certain embodiments, an antisense compound is targeted tonucleotides 9209-9235 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 18, 211, 212, 213 or 214.In certain such embodiments, an antisense compound targeted tonucleotides 9209-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159,399881, 406022, 406023, 406024 or 410732.

In certain embodiments, a target region is nucleotides 10252-10271 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 10252-10271 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 148. In certain suchembodiments, an antisense compound targeted to nucleotides 10252-10271of SEQ ID NO: 2 is selected from ISIS NOs: 399849 or 400005.

In certain embodiments, a target region is nucleotides 10633-10652 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 10633-10652 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 127. In certain suchembodiments, an antisense compound targeted to nucleotides 10633-10652of SEQ ID NO: 2 is selected from ISIS NOs: 395222 or 399944.

In certain embodiments, a target region is nucleotides 11308-11491 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 11308-11491 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 79 or 84. In certain suchembodiments, an antisense compound targeted to nucleotides 11308-11491of SEQ ID NO: 2 is selected from ISIS NOs: 395223, 399850, 399945 or400006.

In certain embodiments, a target region is nucleotides 12715-12734 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 12715-12734 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 88. In certain suchembodiments, an antisense compound targeted to nucleotides 12715-12734of SEQ ID NO: 2 is selected from ISIS NOs: 399851 or 400007.

In certain embodiments, a target region is nucleotides 12928-12947 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 12928-12947 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 111. In certain suchembodiments, an antisense compound targeted to nucleotides 12928-12947of SEQ ID NO: 2 is selected from ISIS NOs: 399852 or 400008.

In certain embodiments, a target region is nucleotides 13681-13700 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 13681-13700 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 85. In certain suchembodiments, an antisense compound targeted to nucleotides 13681-13700of SEQ ID NO: 2 is selected from ISIS NOs: 395201 or 399923.

In certain embodiments, a target region is nucleotides 13746-13779 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 13746-13779 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 100 or 116. In certainsuch embodiments, an antisense compound targeted to nucleotides13746-13779 of SEQ ID NO: 2 is selected from ISIS NOs: 399827, 399828,399983 or 399984.

In certain embodiments, a target region is nucleotides 13816-13847 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 13816-13847 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 117 or 129. In certainsuch embodiments, an antisense compound targeted to nucleotides13816-13847 of SEQ ID NO: 2 is selected from ISIS NOs: 395202, 399829,399924 or 399985.

In certain embodiments, a target region is nucleotides 13903-13945 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 13903-13945 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain suchembodiments, an antisense compound targeted to nucleotides 13903-13945of SEQ ID NO: 2 is selected from ISIS NOs: 395203, 399830, 399925 or399986.

In certain embodiments, a target region is nucleotides 13977-14141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 13977-14141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 83, 136, 137, 140 or 152.In certain such embodiments, an antisense compound targeted tonucleotides 13977-14141 of SEQ ID NO: 2 is selected from ISIS NOs:395204, 395205, 395206, 399831, 399832, 399926, 399927, 399928, 399987or 399988.

In certain embodiments, a target region is nucleotides 14179-14198 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14179-14198 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 132. In certain suchembodiments, an antisense compound targeted to nucleotides 14179-14198of SEQ ID NO: 2 is selected from ISIS NOs: 395207 or 399929.

In certain embodiments, a target region is nucleotides 14267-14286 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14267-14286 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 139. In certain suchembodiments, an antisense compound targeted to nucleotides 14267-14286of SEQ ID NO: 2 is selected from ISIS NOs: 395208 or 399930.

In certain embodiments, a target region is nucleotides 14397-14423 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14397-14423 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain suchembodiments, an antisense compound targeted to nucleotides 14397-14423of SEQ ID NO: 2 is selected from ISIS NOs: 395209, 399833, 399931 or399989.

In certain embodiments, a target region is nucleotides 14441-14460 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14441-14460 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 113. In certain suchembodiments, an antisense compound targeted to nucleotides 14441-14460of SEQ ID NO: 2 is selected from ISIS NOs: 395210 or 399932.

In certain embodiments, a target region is nucleotides 14494-14513 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14494-14513 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 138. In certain suchembodiments, an antisense compound targeted to nucleotides 14494-14513of SEQ ID NO: 2 is selected from ISIS NOs: 395211 or 399933.

In certain embodiments, a target region is nucleotides 14494-14543 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14494-14543 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain suchembodiments, an antisense compound targeted to nucleotides 14494-14543of SEQ ID NO: 2 is selected from ISIS NOs: 395211, 395212, 399933 or399934.

In certain embodiments, a target region is nucleotides 14524-14543 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14524-14543 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 98. In certain suchembodiments, an antisense compound targeted to nucleotides 14524-14543of SEQ ID NO: 2 is selected from ISIS NOs: 395212 or 399934.

In certain embodiments, a target region is nucleotides 14601-14650 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14601-14650 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain suchembodiments, an antisense compound targeted to nucleotides 14601-14650of SEQ ID NO: 2 is selected from ISIS NOs: 395213, 395214, 399935 or399936.

In certain embodiments, a target region is nucleotides 14670-14700 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14670-14700 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 82, 103 or 133. In certainsuch embodiments, an antisense compound targeted to nucleotides14670-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216,399834, 399937, 399938 or 399990.

In certain embodiments, a target region is nucleotides 14675-14700 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14675-14700 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 103 or 133. In certainsuch embodiments, an antisense compound targeted to nucleotides14675-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216,399937 or 399938.

In certain embodiments, a target region is nucleotides 14801-14828 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14801-14828 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 155 or 156. In certainsuch embodiments, an antisense compound targeted to nucleotides14801-14828 of SEQ ID NO: 2 is selected from ISIS NOs: 395217, 399835,399939 or 399991.

In certain embodiments, a target region is nucleotides 14877-14912 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14877-14912 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 215, 216 or 217. Incertain such embodiments, an antisense compound targeted to nucleotides14877-14912 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 399882,406025, 406026 or 410753.

In certain embodiments, a target region is nucleotides 14877-14915 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14877-14915 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217.In certain such embodiments, an antisense compound targeted tonucleotides 14877-14915 of SEQ ID NO: 2 is selected from ISIS NOs:395160, 399797, 399882, 399953, 406025, 406026 or 410753.

In certain embodiments, a target region is nucleotides 14877-14973 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14877-14973 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24,215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228,229, 230, 231, 232, or 233. In certain such embodiments, an antisensecompound targeted to nucleotides 14877-14973 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395160, 395161, 395162, 399797, 399798, 399799, 399882,399883, 399884, 399953, 399954, 399955, 405869, 405870, 405871, 405872,405873, 405874, 405875, 405876, 406025, 406026, 406027, 406028, 406029,406030, 406031, 406032, 410733, 410753, or 410754.

In certain embodiments, a target region is nucleotides 14916-14943 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14916-14943 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220,221, 222 or 223. In certain such embodiments, an antisense compoundtargeted to nucleotides 14916-14943 of SEQ ID NO: 2 is selected fromISIS NOs: 395161, 399798, 399883, 399954, 405869, 405870, 405871,405872, 405873 or 405874.

In certain embodiments, a target region is nucleotides 14916-14973 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14916-14973 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219,220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 223.In certain such embodiments, an antisense compound targeted tonucleotides 14916-14973 of SEQ ID NO: 2 is selected from ISIS NOs:395161, 395162, 399798, 399799, 399883, 399884, 399954, 399955, 405869,405870, 405871, 405872, 405873, 405874, 405875, 405876, 406027, 406028,406029, 406030, 406031, 406032, 410733, or 410754.

In certain embodiments, a target region is nucleotides 14925-14951 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14925-14951 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. Incertain such embodiments, an antisense compound targeted to nucleotides14925-14951 of SEQ ID NO: 2 is selected from ISIS NOs: 405875, 405876,406027 or 406028.

In certain embodiments, a target region is nucleotides 14934-14963 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14934-14963 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, 231 or232. In certain such embodiments, an antisense compound targeted tonucleotides 14934-14963 of SEQ ID NO: 2 is selected from ISIS NOs:399799, 399955, 406029, 406030, 406031, 406032 or 410733.

In certain embodiments, a target region is nucleotides 14946-14973 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14946-14973 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain suchembodiments, an antisense compound targeted to nucleotides 14946-14973of SEQ ID NO: 2 is selected from ISIS NOs: 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 14979-14998 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 14979-14998 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 25. In certain suchembodiments, an antisense compound targeted to nucleotides 14979-14998of SEQ ID NO: 2 is selected from ISIS NOs: 395163 or 399885.

In certain embodiments, a target region is nucleotides 15254-15280 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15254-15280 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 235, 236 or 237. Incertain such embodiments, an antisense compound targeted to nucleotides15254-15280 of SEQ ID NO: 2 is selected from ISIS NOs: 405526, 405604,406033 or 410756.

In certain embodiments, a target region is nucleotides 15254-15328 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15254-15328 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237,238, 239, 240, 241, 242, 243, 243, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, or 447. In certain such embodiments,an antisense compound targeted to nucleotides 15254-15328 of SEQ ID NO:2 is selected from ISIS NOs: 395164, 395165, 399800, 399886, 399887,399956, 405526, 405604, 405877, 405878, 405879, 405880, 405881, 405882,405883, 405884, 406033, 406034, 406035, 406036, 406037, 406038, 409126,410536, 410537, 410538, 410539, 410583, 410584, 410585, 410586, 410587,410588, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410658,410659, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667,410668, 410669, 410670, 410671, 410756, 410757, or 410758.

In certain embodiments, a target region is nucleotides 15264-15290 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15264-15290 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 27, 238 or 239. Incertain such embodiments, an antisense compound targeted to nucleotides15264-15290 of SEQ ID NO: 2 is selected from ISIS NOs: 395164, 399800,399886, 399956, 406034, or 406035.

In certain embodiments, a target region is nucleotides 15279-15305 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15279-15305 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 240, 241 or 242. Incertain such embodiments, an antisense compound targeted to nucleotides15279-15305 of SEQ ID NO: 2 is selected from ISIS NOs: 406036, 406037,or 410757.

In certain embodiments, a target region is nucleotides 15291-15318 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15291-15318 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 243, 244, 245, 246,247, 248, 249, 250, or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 15291-15318 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,406038, 409126, 410536, 410537, 410583, 410584, 410585, 410586, 410587,410588, 410589, 410590, 410658, 410659, 410660, 410661, 410662, 410663,410664, 410665, or 410666.

In certain embodiments, a target region is nucleotides 15292-15319 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15292-15319 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247,248, 249, 250 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 15292-15319 of SEQ ID NO: 2 is selected fromISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,406038, 409126, 410537, 410584, 410585, 410586, 410587, 410588, 410589,410590, 410659, 410660, 410661, 410662, 410663, 410664, 410665, or410666.

In certain embodiments, a target region is nucleotides 15293-15320 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15293-15320 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248,249, 250, 251, 252 or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 15293-15320 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 405883, 406038, 409126, 410585, 410586, 410587, 410588, 410589,410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665,410666, 410667, or 410668.

In certain embodiments, a target region is nucleotides 15294-15321 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249,250, 251, 252, 253 or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590,410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666,410667, 410668 or 410669.

In certain embodiments, a target region is nucleotides 15294-15321 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249,250, 251, 252, 253 or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590,410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666,410667, 410668 or 410669.

In certain embodiments, a target region is nucleotides 15295-15322 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15295-15322 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250,250, 251, 252, 253, 254 or 447. In certain such embodiments, anantisense compound targeted to nucleotides 15295-15322 of SEQ ID NO: 2is selected from ISIS NOs: 395165, 399887, 405878, 405879, 405880,405881, 405882, 405883, 405884, 409126, 410538, 410587, 410588, 410589,410590, 410591, 410592, 410593, 410594, 410662, 410663, 410664, 410665,410666, 410667, 410668, 410669 or 410670.

In certain embodiments, a target region is nucleotides 15296-15315 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15296-15315 of SEQ ID NO: 2. In one such embodiment, anantisense compound targeted to nucleotides 15296-15315 of SEQ ID NO: 2is ISIS NO: 405879.

In certain embodiments, a target region is nucleotides 15296-15323 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15296-15323 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251,252, 253, 254, 255 or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 15296-15323 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395165, 399887, 405879, 405880, 405881, 405882, 405883,405884, 409126, 410538, 410539, 410588, 410589, 410590, 410591, 410592,410593, 410594, 410595, 410663, 410664, 410665, 410666, 410667, 410668,410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15297-15323 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15297-15323 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252,253, 254, 255 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 15297-15323 of SEQ ID NO: 2 is selected fromISIS NOs: 395165, 399887, 405880, 405881, 405882, 405883, 405884,409126, 410538, 410539, 410589, 410590, 410591, 410592, 410593, 410594,410595, 410664, 410665, 410666, 410667, 410668, 410669, 410670 or410671.

In certain embodiments, a target region is nucleotides 15298-15323 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15298-15323 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253,254, 255 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 15298-15323 of SEQ ID NO: 2 is selected fromISIS NOs: 395165, 399887, 405881, 405882, 405883, 405884, 409126,410538, 410539, 410590, 410591, 410592, 410593, 410594, 410595, 410665,410666, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15299-15323 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15299-15323 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254 or255. In certain such embodiments, an antisense compound targeted tonucleotides 15299-15323 of SEQ ID NO: 2 is selected from ISIS NOs:405881, 405882, 405883, 405884, 410538, 410539, 410590, 410591, 410592,410593, 410594, 410595, 410666, 410667, 410668, 410669, 410670 or410671.

In certain embodiments, a target region is nucleotides 15300-15323 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15300-15323 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254 or 255.In certain such embodiments, an antisense compound targeted tonucleotides 15300-15323 of SEQ ID NO: 2 is selected from ISIS NOs:405882, 405883, 405884, 410538, 410539, 410591, 410592, 410593, 410594,410595, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15301-15328 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15301-15328 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255 or 256.In certain such embodiments, an antisense compound targeted tonucleotides 15301-15328 of SEQ ID NO: 2 is selected from ISIS NOs:405883, 405884, 410538, 410539, 410592, 410593, 410594, 410595, 410668,410669, 410670, 410671 or 410758.

In certain embodiments, a target region is nucleotides 15330-15355 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15330-15355 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 29, 257, 258 or 259. Incertain such embodiments, an antisense compound targeted to nucleotides15330-15355 of SEQ ID NO: 2 is selected from ISIS NOs: 395166, 399888,406039, 406040 or 406041.

In certain embodiments, a target region is nucleotides 15330-15490 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15330-15490 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 29, 30, 86, 257, 258, 259,260, 261, 262 or 263. In certain such embodiments, an antisense compoundtargeted to nucleotides 15330-15490 of SEQ ID NO: 2 is selected fromISIS NOs: 395166, 399801, 399853, 399888, 399957, 400009, 406039,406040, 406041, 406042, 406043, 406044 or 410759.

In certain embodiments, a target region is nucleotides 15339-15366 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15339-15366 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 30, 260, 261 or 262. Incertain such embodiments, an antisense compound targeted to nucleotides15339-15366 of SEQ ID NO: 2 is selected from ISIS NOs: 399801, 399957,406042, 406043 or 406044.

In certain embodiments, a target region is nucleotides 15358-15490 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 15358-15490 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 86 or 263. In certain suchembodiments, an antisense compound targeted to nucleotides 15358-15490of SEQ ID NO: 2 is selected from ISIS NOs: 399853, 400009 or 410759.

In certain embodiments, a target region is nucleotides 16134-16153 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 16134-16153 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 97. In certain suchembodiments, an antisense compound targeted to nucleotides 16134-16153of SEQ ID NO: 2 is selected from ISIS NOs: 399854 or 400010.

In certain embodiments, a target region is nucleotides 16668-16687 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 16668-16687 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 120. In certain suchembodiments, an antisense compound targeted to nucleotides 16668-16687of SEQ ID NO: 2 is selected from ISIS NOs: 399855 or 400011.

In certain embodiments, a target region is nucleotides 17267-17286 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 17267-17286 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 150. In certain suchembodiments, an antisense compound targeted to nucleotides 17267-17286of SEQ ID NO: 2 is selected from ISIS NOs: 399856 or 400012.

In certain embodiments, a target region is nucleotides 18377-18427 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18377-18427 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 115 or 134. In certainsuch embodiments, an antisense compound targeted to nucleotides18377-18427 of SEQ ID NO: 2 is selected from ISIS NOs: 399857, 399858,400013 or 400014.

In certain embodiments, a target region is nucleotides 18561-18580 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18561-18580 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 157. In certain suchembodiments, an antisense compound targeted to nucleotides 18561-18580of SEQ ID NO: 2 is selected from ISIS NOs: 395224 or 399946.

In certain embodiments, a target region is nucleotides 18591-18618 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18591-18618 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268 or 269.In certain such embodiments, an antisense compound targeted tonucleotides 18591-18618 of SEQ ID NO: 2 is selected from ISIS NOs:395168, 399890, 405909, 405910, 405911 or 406045.

In certain embodiments, a target region is nucleotides 18591-18646 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18591-18646 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 32, 32, 266, 267, 268,269, 270, 271 or 272. In certain such embodiments, an antisense compoundtargeted to nucleotides 18591-18646 of SEQ ID NO: 2 is selected fromISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045, 410761or 410762.

In certain embodiments, a target region is nucleotides 18591-18668 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18591-18668 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, 269,270, 271, 272 or 273. In certain such embodiments, an antisense compoundtargeted to nucleotides 18591-18668 of SEQ ID NO: 2 is selected fromISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045,410761, 410762 or 410763.

In certain embodiments, a target region is nucleotides 18695-18746 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18695-18746 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284 or 285. In certain such embodiments,an antisense compound targeted to nucleotides 18695-18746 of SEQ ID NO:2 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915,405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734 or410764.

In certain embodiments, a target region is nucleotides 18705-18730 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18705-18730 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 275, 276 or 277. Incertain such embodiments, an antisense compound targeted to nucleotides18705-18730 of SEQ ID NO: 2 is selected from ISIS NOs: 395169, 399891,405913, 405914 or 405915.

In certain embodiments, a target region is nucleotides 18709-18736 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18709-18736 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 276, 277, 278, 279 or 280.In certain such embodiments, an antisense compound targeted tonucleotides 18709-18736 of SEQ ID NO: 2 is selected from ISIS NOs:405914, 405915, 405916, 405917 or 410734.

In certain embodiments, a target region is nucleotides 18719-18746 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 18719-18746 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 281, 282, 283, 284 or 285.In certain such embodiments, an antisense compound targeted tonucleotides 18719-18746 of SEQ ID NO: 2 is selected from ISIS NOs:405918, 405919, 405920, 405921 or 405922.

In certain embodiments, a target region is nucleotides 19203-20080 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19203-20080 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39,40, 41, 105, 128, 149, 151, 288, 289, 290, 291, 292, 293, 294, 295, 296,297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310,311, 312, 313, 314, 315, 316, 317 or 318. In certain such embodiments,an antisense compound targeted to nucleotides 19203-20080 of SEQ ID NO:2 is selected from ISIS NOs: 395170, 395171, 395172, 395173, 395174,395175, 399802, 399803, 399804, 399805, 399859, 399860, 399892, 399893,399894, 399895, 399896, 399897, 399958, 399959, 399960, 399961, 400015,400016, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930,405931, 405932, 405933, 405934, 405935, 405936, 405937, 405938, 405939,405940, 405941, 405942, 405943, 405944, 405945, 405946, 405947, 410735,410736, 410737, 410767, 410768 or 410769.

In certain embodiments, a target region is nucleotides 19931-19952 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19931-19952 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 149 or 188. In certainsuch embodiments, an antisense compound targeted to nucleotides19931-19952 of SEQ ID NO: 2 is selected from ISIS NOs: 395170, 399892 or405923.

In certain embodiments, a target region is nucleotides 19954-19981 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19954-19981 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293.In certain such embodiments, an antisense compound targeted tonucleotides 19954-19981 of SEQ ID NO: 2 is selected from ISIS NOs:395171, 399893, 405924, 405925, 405926 or 410735.

In certain embodiments, a target region is nucleotides 19964-19990 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19964-19990 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 35, 128, 128, 294 or 295.In certain such embodiments, an antisense compound targeted tonucleotides 19964-19990 of SEQ ID NO: 2 is selected from ISIS NOs:395172, 399802, 399894, 399958, 405927 or 405928.

In certain embodiments, a target region is nucleotides 19973-19999 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19973-19999 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 36, 296, 297 or 298. Incertain such embodiments, an antisense compound targeted to nucleotides19973-19999 of SEQ ID NO: 2 is selected from ISIS NOs: 395173, 399895,405929, 405930 or 405931.

In certain embodiments, a target region is nucleotides 19982-20009 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19982-20009 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 37, 299, 300, 301 or 302.In certain such embodiments, an antisense compound targeted tonucleotides 19982-20009 of SEQ ID NO: 2 is selected from ISIS NOs:399803, 399959, 405932, 405933, 405934 or 405935.

In certain embodiments, a target region is nucleotides 19992-20016 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 19992-20016 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 38, 303 or 304. In certainsuch embodiments, an antisense compound targeted to nucleotides19992-20016 of SEQ ID NO: 2 is selected from ISIS NOs: 395174, 399896,405936 or 410736.

In certain embodiments, a target region is nucleotides 20016-20042 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20016-20042 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 305 or 306. In certainsuch embodiments, an antisense compound targeted to nucleotides20016-20042 of SEQ ID NO: 2 is selected from ISIS NOs: 405937 or 410768.

In certain embodiments, a target region is nucleotides 20025-20052 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20025-20052 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310.In certain such embodiments, an antisense compound targeted tonucleotides 20025-20052 of SEQ ID NO: 2 is selected from ISIS NOs:395175, 399897, 405938, 405939, 405940 or 405941.

In certain embodiments, a target region is nucleotides 20036-20062 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20036-20062 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313 or 314.In certain such embodiments, an antisense compound targeted tonucleotides 20036-20062 of SEQ ID NO: 2 is selected from ISIS NOs:399804, 399960, 405942, 405943, 405944 or 410737.

In certain embodiments, a target region is nucleotides 20045-20070 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20045-20070 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 41, 315, 316 or 317. Incertain such embodiments, an antisense compound targeted to nucleotides20045-20070 of SEQ ID NO: 2 is selected from ISIS NOs: 399805, 399961,405945, 405946 or 405947.

In certain embodiments, a target region is nucleotides 20100-20119 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20100-20119 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 158. In certain suchembodiments, an antisense compound targeted to nucleotides 20100-20119of SEQ ID NO: 2 is selected from ISIS NOs: 399861 or 400017.

In certain embodiments, a target region is nucleotides 20188-20207 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20188-20207 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 109. In certain suchembodiments, an antisense compound targeted to nucleotides 20188-20207of SEQ ID NO: 2 is selected from ISIS NOs: 399862 or 400018.

In certain embodiments, a target region is nucleotides 20624-20650 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20624-20650 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 141, 320 or 321. Incertain such embodiments, an antisense compound targeted to nucleotides20624-20650 of SEQ ID NO: 2 is selected from ISIS NOs: 399863, 400019,405949 or 405950.

In certain embodiments, a target region is nucleotides 20624-20759 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20624-20759 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48,49, 50, 51, 87, 141, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329,330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343,344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357,358 or 359. In certain such embodiments, an antisense compound targetedto nucleotides 20624-20759 of SEQ ID NO: 2 is selected from ISIS NOs:395177, 395178, 395179, 399807, 399808, 399809, 399810, 399811, 399812,399813, 399863, 399899, 399900, 399901, 399963, 399964, 399965, 399966,399967, 399968, 399969, 400019, 405557, 405885, 405886, 405887, 405888,405889, 405890, 405891, 405892, 405949, 405950, 405951, 405952, 405953,405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962,405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971,405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599,410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675,410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739,410740 or 410770.

In certain embodiments, a target region is nucleotides 20629-20804 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20629-20804 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48,49, 50, 51, 87, 119, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329,330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343,344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 356,357, 358, 359 or 360. In certain such embodiments, an antisense compoundtargeted to nucleotides 20629-20804 of SEQ ID NO: 2 is selected fromISIS NOs: 395177, 395178, 395179, 395180, 399807, 399808, 399809,399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399963,399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886,405887, 405888, 405889, 405890, 405891, 405892, 405949, 405950, 405951,405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960,405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969,405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597,410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673,410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682,410738, 410739, 410740, 410770 or 410771.

In certain embodiments, a target region is nucleotides 20633-20660 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20633-20660 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 322, 323, 324 or 325.In certain such embodiments, an antisense compound targeted tonucleotides 20633-20660 of SEQ ID NO: 2 is selected from ISIS NOs:399807, 399963, 405951, 405952, 405953 or 410738.

In certain embodiments, a target region is nucleotides 20635-20781 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20635-20781 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48,49, 50, 51, 87, 119, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332,333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346,347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. Incertain such embodiments, an antisense compound targeted to nucleotides20635-20781 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178,395179, 395180, 399807, 399808, 399809, 399810, 399811, 399812, 399813,399899, 399900, 399901, 399902, 399963, 399964, 399965, 399966, 399967,399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890,405891, 405892, 405952, 405953, 405954, 405955, 405956, 405957, 405958,405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967,405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540,410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604,410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680,410681, 410682, 410738, 410739, 410740 or 410770.

In certain embodiments, a target region is nucleotides 20643-20662 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20643-20662 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 326. In certain suchembodiments, an antisense compound targeted to nucleotides 20643-20662of SEQ ID NO: 2 is selected from ISIS NOs: 405954.

In certain embodiments, a target region is nucleotides 20657-20676 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20657-20676 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 327. In certain suchembodiments, an antisense compound targeted to nucleotides 20657-20676of SEQ ID NO: 2 is selected from ISIS NOs: 410770.

In certain embodiments, a target region is nucleotides 20670-20697 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20670-20697 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331 or 332.In certain such embodiments, an antisense compound targeted tonucleotides 20670-20697 of SEQ ID NO: 2 is selected from ISIS NOs:405955, 405956, 405957, 405958 or 405959.

In certain embodiments, a target region is nucleotides 20680-20706 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20680-20706 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335 or 336.In certain such embodiments, an antisense compound targeted tonucleotides 20680-20706 of SEQ ID NO: 2 is selected from ISIS NOs:399808, 399964, 405960, 405961, 405962 or 410739.

In certain embodiments, a target region is nucleotides 20683-20781 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20683-20781 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49,50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345,346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359.In certain such embodiments, an antisense compound targeted tonucleotides 20683-20781 of SEQ ID NO: 2 is selected from ISIS NOs:395177, 395178, 395179, 395180, 399808, 399809, 399810, 399811, 399812,399813, 399899, 399900, 399901, 399902, 399964, 399965, 399966, 399967,399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890,405891, 405892, 405962, 405963, 405964, 405965, 405966, 405967, 405968,405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596,410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672,410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681,410682, 410739 or 410740.

In certain embodiments, a target region is nucleotides 20689-20715 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20689-20715 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 45, 46, 337 or 338. Incertain such embodiments, an antisense compound targeted to nucleotides20689-20715 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 399809,399899, 399965, 405963 or 405964.

In certain embodiments, a target region is nucleotides 20698-20725 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20698-20725 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 47, 47, 339, 340, 341 or342. In certain such embodiments, an antisense compound targeted tonucleotides 20698-20725 of SEQ ID NO: 2 is selected from ISIS NOs:399810, 399966, 405965, 405966, 405967 or 405968.

In certain embodiments, a target region is nucleotides 20709-20736 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20709-20736 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 48, 48, 343, 344, 345 or346. In certain such embodiments, an antisense compound targeted tonucleotides 20709-20736 of SEQ ID NO: 2 is selected from ISIS NOs:399811, 399967, 405885, 405969, 405970 or 410740.

In certain embodiments, a target region is nucleotides 20717-20744 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20717-20744 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349,350, 351, 352 or 353. In certain such embodiments, an antisense compoundtargeted to nucleotides 20717-20744 of SEQ ID NO: 2 is selected fromISIS NOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889,405890, 405891, 405892, 410596, 410597, 410598, 410599, 410600, 410601,410672, 410673, 410674, 410675, 410676, 410677 or 410678.

In certain embodiments, a target region is nucleotides 20718-20745 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20718-20745 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 347, 348, 349,350, 351, 352 or 353. In certain such embodiments, an antisense compoundtargeted to nucleotides 20718-20745 of SEQ ID NO: 2 is selected fromISIS NOs: 395178, 399812, 399900, 399968, 405557, 405886, 405887,405888, 405889, 405890, 405891, 405892, 410596, 410597, 410598, 410599,410600, 410601, 410672, 410673, 410674, 410675, 410676, 410677, 410678or 410679.

In certain embodiments, a target region is nucleotides 20719-20746 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20719-20746 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 348, 349, 350,351, 352, 353 or 354. In certain such embodiments, an antisense compoundtargeted to nucleotides 20719-20746 of SEQ ID NO: 2 is selected fromISIS NOs: 395178, 399812, 399900, 399968, 405557, 405887, 405888,405889, 405890, 405891, 405892, 408653, 410596, 410597, 410598, 410599,410600, 410601, 410602, 410672, 410673, 410674, 410675, 410676, 410677,410678, 410679 or 410680.

In certain embodiments, a target region is nucleotides 20720-20747 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20720-20747 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351,352, 353, 354 or 355. In certain such embodiments, an antisense compoundtargeted to nucleotides 20720-20747 of SEQ ID NO: 2 is selected fromISIS NOs: 395178, 399812, 399900, 399968, 405557, 405888, 405889,405890, 405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600,410601, 410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678,410679, 410680 or 410681.

In certain embodiments, a target region is nucleotides 20721-20748 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20721-20748 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352,353, 354, 355 or 356. In certain such embodiments, an antisense compoundtargeted to nucleotides 20721-20748 of SEQ ID NO: 2 is selected fromISIS NOs: 395178, 399812, 399900, 399968, 405557, 405889, 405890,405891, 405892, 405971, 408653, 410540, 410598, 410599, 410600, 410601,410602, 410603, 410604, 410674, 410675, 410676, 410677, 410678, 410679,410680, 410681 or 410682.

In certain embodiments, a target region is nucleotides 20722-20749 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20722-20749 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353,354, 355, 356 or 357. In certain such embodiments, an antisense compoundtargeted to nucleotides 20722-20749 of SEQ ID NO: 2 is selected fromISIS NOs: 395178, 399900, 405557, 405889, 405890, 405891, 405892,405971, 405972, 408653, 410540, 410598, 410599, 410600, 410601, 410602,410603, 410604, 410675, 410676, 410677, 410678, 410679, 410680, 410681or 410682.

In certain embodiments, a target region is nucleotides 20727-20752 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20727-20752 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 87, 354, 355, 356 or 357.In certain such embodiments, an antisense compound targeted tonucleotides 20727-20752 of SEQ ID NO: 2 is selected from ISIS NOs:395179, 399901, 405971, 405972, 408653, 410540, 410602, 410603, 410604,410680, 410681 or 410682.

In certain embodiments, a target region is nucleotides 20735-20759 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20735-20759 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 51, 358 or 359. In certainsuch embodiments, an antisense compound targeted to nucleotides20735-20759 of SEQ ID NO: 2 is selected from ISIS NOs: 399813, 399969,405973 or 405974.

In certain embodiments, a target region is nucleotides 20762-21014 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20762-21014 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 93, 119 or 360. In certainsuch embodiments, an antisense compound targeted to nucleotides20762-21014 of SEQ ID NO: 2 is selected from ISIS NOs: 395180, 399864,399902, 400020 or 410771.

In certain embodiments, a target region is nucleotides 20785-21014 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 20785-21014 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 93 or 360. In certain suchembodiments, an antisense compound targeted to nucleotides 20785-21014of SEQ ID NO: 2 is selected from ISIS NOs: 399864, 400020 or 410771.

In certain embodiments, a target region is nucleotides 21082-21107 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21082-21107 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 95 or 361. In certain suchembodiments, an antisense compound targeted to nucleotides 21082-21107of SEQ ID NO: 2 is selected from ISIS NOs: 399865, 400021 or 405975.

In certain embodiments, a target region is nucleotides 21082-21152 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21082-21152 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 53, 54, 95, 361, 362, 363,364, 365, 366, 367, 368, 369, 370 or 371. In certain such embodiments,an antisense compound targeted to nucleotides 21082-21152 of SEQ ID NO:2 is selected from ISIS NOs: 395182, 399814, 399865, 399904, 399970,400021, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982,405983, 410741 or 410772.

In certain embodiments, a target region is nucleotides 21091-21114 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21091-21114 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 53, 362 or 363. In certainsuch embodiments, an antisense compound targeted to nucleotides21091-21114 of SEQ ID NO: 2 is selected from ISIS NOs: 399814, 399970,405976 or 405977.

In certain embodiments, a target region is nucleotides 21118-21144 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21118-21144 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 54, 365, 366 or 367. Incertain such embodiments, an antisense compound targeted to nucleotides21118-21144 of SEQ ID NO: 2 is selected from ISIS NOs: 395182, 399904,405978, 405979 or 405980.

In certain embodiments, a target region is nucleotides 21127-21152 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21127-21152 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 368, 369, 370 or 371. Incertain such embodiments, an antisense compound targeted to nucleotides21127-21152 of SEQ ID NO: 2 is selected from ISIS NOs: 405981, 405982,405983 or 410741.

In certain embodiments, a target region is nucleotides 21181-21209 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21181-21209 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 372, 373, 374 or 375.In certain such embodiments, an antisense compound targeted tonucleotides 21181-21209 of SEQ ID NO: 2 is selected from ISIS NOs:399815, 399971, 405564, 405641, 405984, 405985, 405986 or 405987.

In certain embodiments, a target region is nucleotides 21181-21211 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21181-21211 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374 or375. In certain such embodiments, an antisense compound targeted tonucleotides 21181-21211 of SEQ ID NO: 2 is selected from ISIS NOs:399815, 399816, 399971, 399972, 405564, 405641, 405984, 405985, 405986or 405987.

In certain embodiments, a target region is nucleotides 21183-21211 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21183-21211 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 56, 373, 374 or 375.In certain such embodiments, an antisense compound targeted tonucleotides 21183-21211 of SEQ ID NO: 2 is selected from ISIS NOs:399815, 399816, 399971, 399972, 405564, 405641, 405985, 405986 or405987.

In certain embodiments, a target region is nucleotides 21481-21500 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21481-21500 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 143. In certain suchembodiments, an antisense compound targeted to nucleotides 21481-21500of SEQ ID NO: 2 is selected from ISIS NOs: 399866 or 400022.

In certain embodiments, a target region is nucleotides 21589-21608 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21589-21608 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 89. In certain suchembodiments, an antisense compound targeted to nucleotides 21589-21608of SEQ ID NO: 2 is selected from ISIS NOs: 399867 or 400023.

In certain embodiments, a target region is nucleotides 21692-21719 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 21692-21719 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 123, 448, 449, 450, 451,452, 453, 454 or 455. In certain such embodiments, an antisense compoundtargeted to nucleotides 21692-21719 of SEQ ID NO: 2 is selected fromISIS NOs: 399868, 400024, 406478, 406479, 406480, 406481, 406482,406483, 406484 or 406485.

In certain embodiments, a target region is nucleotides 22000-22227 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22000-22227 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392,393, 394, 395 or 396. In certain such embodiments, an antisense compoundtargeted to nucleotides 22000-22227 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552,410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621,410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701,410702 or 410703.

In certain embodiments, a target region is nucleotides 22096-22115 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22096-22115 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 57. In certain suchembodiments, an antisense compound targeted to nucleotides 22096-22115of SEQ ID NO: 2 is selected from ISIS NOs: 395183 or 399905.

In certain embodiments, a target region is nucleotides 22096-22223 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22096-22223 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 376, 377, 378,379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391 or 392.In certain such embodiments, an antisense compound targeted tonucleotides 22096-22223 of SEQ ID NO: 2 is selected from ISIS NOs:395183, 395184, 395185, 399905, 399906, 399907, 405988, 405989, 405990,410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410549,410550, 410551, 410552, 410553, 410605, 410606, 410607, 410608, 410609,410610, 410611, 410612, 410613, 410614, 410615, 410616, 410617, 410618,410619, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690,410691, 410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699or 410773.

In certain embodiments, a target region is nucleotides 22096-22311 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22096-22311 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 126, 126, 376,377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390,391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 401, 401, 402,402, 402, 403, 403, 403, 404, 404, 404, 405, 405, 405 or 406. In certainsuch embodiments, an antisense compound targeted to nucleotides22096-22311 of SEQ ID NO: 2 is selected from ISIS NOs: 395183, 395184,395185, 395225, 399905, 399906, 399907, 399947, 405988, 405989, 405990,405991, 405992, 405993, 405994, 405995, 410541, 410542, 410543, 410544,410545, 410546, 410547, 410548, 410549, 410550, 410551, 410552, 410553,410554, 410555, 410556, 410557, 410558, 410559, 410560, 410561, 410605,410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614,410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623,410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632,410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691,410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699, 410700,410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709,410710, 410711, 410712, 410773 or 410774.

In certain embodiments, a target region is nucleotides 22133-22160 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22133-22160 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380,381, 382, 383 or 384. In certain such embodiments, an antisense compoundtargeted to nucleotides 22133-22160 of SEQ ID NO: 2 is selected fromISIS NOs: 405988, 405989, 410541, 410542, 410543, 410544, 410545,410546, 410547, 410605, 410606, 410607, 410608, 410609, 410610, 410611,410612, 410613, 410683, 410684, 410685, 410686, 410687, 410688, 410689,410690 or 410691.

In certain embodiments, a target region is nucleotides 22133-22163 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22133-22163 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379,380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, anantisense compound targeted to nucleotides 22133-22163 of SEQ ID NO: 2is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990,410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410605,410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614,410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691,410692 or 410693.

In certain embodiments, a target region is nucleotides 22134-22161 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22134-22161 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380,381, 382, 383 or 384. In certain such embodiments, an antisense compoundtargeted to nucleotides 22134-22161 of SEQ ID NO: 2 is selected fromISIS NOs: 395184, 399906, 405988, 405989, 410542, 410543, 410544,410545, 410546, 410547, 410606, 410607, 410608, 410609, 410610, 410611,410612, 410613, 410684, 410685, 410686, 410687, 410688, 410689, 410690,410691 or 410692.

In certain embodiments, a target region is nucleotides 22135-22162 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22135-22162 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381,382, 383, 384 or 385. In certain such embodiments, an antisense compoundtargeted to nucleotides 22135-22162 of SEQ ID NO: 2 is selected fromISIS NOs: 395184, 399906, 405988, 405989, 410543, 410544, 410545,410546, 410547, 410548, 410607, 410608, 410609, 410610, 410611, 410612,410613, 410614, 410685, 410686, 410687, 410688, 410689, 410690, 410691,410692 or 410693.

In certain embodiments, a target region is nucleotides 22136-22163 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22136-22163 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382,383, 384, 385 or 386. In certain such embodiments, an antisense compoundtargeted to nucleotides 22136-22163 of SEQ ID NO: 2 is selected fromISIS NOs: 395184, 399906, 405988, 405989, 405990, 410544, 410545,410546, 410547, 410548, 410608, 410609, 410610, 410611, 410612, 410613,410614, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or410693.

In certain embodiments, a target region is nucleotides 22137-22163 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22137-22163 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383,384, 385 or 386. In certain such embodiments, an antisense compoundtargeted to nucleotides 22137-22163 of SEQ ID NO: 2 is selected fromISIS NOs: 395184, 399906, 405988, 405989, 405990, 410545, 410546,410547, 410548, 410609, 410610, 410611, 410612, 410613, 410614, 410687,410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22138-22163 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22138-22163 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384,385 or 386. In certain such embodiments, an antisense compound targetedto nucleotides 22138-22163 of SEQ ID NO: 2 is selected from ISIS NOs:395184, 399906, 405988, 405989, 405990, 410546, 410547, 410548, 410610,410611, 410612, 410613, 410614, 410688, 410689, 410690, 410691, 410692or 410693.

In certain embodiments, a target region is nucleotides 22189-22239 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22189-22239 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 387, 388, 389, 390,391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404,405 or 406. In certain such embodiments, an antisense compound targetedto nucleotides 22189-22239 of SEQ ID NO: 2 is selected from ISIS NOs:395185, 399907, 405991, 405992, 405993, 405994, 405995, 410549, 410550,410551, 410552, 410553, 410554, 410555, 410556, 410557, 410558, 410559,410560, 410561, 410615, 410616, 410617, 410618, 410619, 410620, 410621,410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630,410631, 410632, 410694, 410695, 410696, 410697, 410698, 410699, 410700,410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709,410710, 410711, 410712, 410773 or 410774.

In certain embodiments, a target region is nucleotides 22199-22226 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22199-22226 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391,392, 393, 394 or 395. In certain such embodiments, an antisense compoundtargeted to nucleotides 22199-22226 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 410549, 410550, 410551, 410552,410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619, 410620,410621, 410622, 410694, 410695, 410696, 410697, 410698, 410699, 410700,410701 or 410702.

In certain embodiments, a target region is nucleotides 22199-22227 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22199-22227 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391,392, 393, 394, 395 or 396. In certain such embodiments, an antisensecompound targeted to nucleotides 22199-22227 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395185, 399907, 405991, 405992, 410549, 410550, 410551,410552, 410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619,410620, 410621, 410622, 410623, 410694, 410695, 410696, 410697, 410698,410699, 410700, 410701, 410702 or 410703.

In certain embodiments, a target region is nucleotides 22200-22227 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22200-22227 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392,393, 394, 395 or 396. In certain such embodiments, an antisense compoundtargeted to nucleotides 22200-22227 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552,410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621,410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701,410702 or 410703.

In certain embodiments, a target region is nucleotides 22201-22228 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22201-22228 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393,394, 395, 396 or 397. In certain such embodiments, an antisense compoundtargeted to nucleotides 22201-22228 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 410551, 410552, 410553,410554, 410555, 410556, 410617, 410618, 410619, 410620, 410621, 410622,410623, 410624, 410696, 410697, 410698, 410699, 410700, 410701, 410702,410703 or 410704.

In certain embodiments, a target region is nucleotides 22202-22229 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22202-22229 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394,395, 396, 397 or 398. In certain such embodiments, an antisense compoundtargeted to nucleotides 22202-22229 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 405993, 410552, 410553,410554, 410555, 410556, 410618, 410619, 410620, 410621, 410622, 410623,410624, 410625, 410697, 410698, 410699, 410700, 410701, 410702, 410703,410704 or 410705.

In certain embodiments, a target region is nucleotides 22203-22230 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22203-22230 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395,396, 397, 398 or 399. In certain such embodiments, an antisense compoundtargeted to nucleotides 22203-22230 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 405993, 410553, 410554,410555, 410556, 410557, 410619, 410620, 410621, 410622, 410623, 410624,410625, 410626, 410698, 410699, 410700, 410701, 410702, 410703, 410704,410705 or 410706.

In certain embodiments, a target region is nucleotides 22204-22231 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22204-22231 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396,397, 398, 399 or 400. In certain such embodiments, an antisense compoundtargeted to nucleotides 22204-22231 of SEQ ID NO: 2 is selected fromISIS NOs: 395185, 399907, 405991, 405992, 405993, 405994, 410554,410555, 410556, 410557, 410620, 410621, 410622, 410623, 410624, 410625,410626, 410627, 410699, 410700, 410701, 410702, 410703, 410704, 410705,410706 or 410707.

In certain embodiments, a target region is nucleotides 22205-22232 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22205-22232 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397,398, 399, 400 or 401. In certain such embodiments, an antisense compoundtargeted to nucleotides 22205-22232 of SEQ ID NO: 2 is selected fromISIS NOs: 405991, 405992, 405993, 405994, 410554, 410555, 410556,410557, 410558, 410620, 410621, 410622, 410623, 410624, 410625, 410626,410627, 410628, 410700, 410701, 410702, 410703, 410704, 410705, 410706,410707 or 410708.

In certain embodiments, a target region is nucleotides 22206-22233 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22206-22233 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398,399, 400, 401 or 402. In certain such embodiments, an antisense compoundtargeted to nucleotides 22206-22233 of SEQ ID NO: 2 is selected fromISIS NOs: 405991, 405992, 405993, 405994, 405995, 410555, 410556,410557, 410558, 410621, 410622, 410623, 410624, 410625, 410626, 410627,410628, 410629, 410701, 410702, 410703, 410704, 410705, 410706, 410707,410708 or 410709.

In certain embodiments, a target region is nucleotides 22207-22234 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22207-22234 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399,400, 401, 402 or 403. In certain such embodiments, an antisense compoundtargeted to nucleotides 22207-22234 of SEQ ID NO: 2 is selected fromISIS NOs: 405992, 405993, 405994, 405995, 410555, 410556, 410557,410558, 410559, 410622, 410623, 410624, 410625, 410626, 410627, 410628,410629, 410630, 410702, 410703, 410704, 410705, 410706, 410707, 410708,410709 or 410710.

In certain embodiments, a target region is nucleotides 22208-22235 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22208-22235 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400,401, 402, 403 or 404. In certain such embodiments, an antisense compoundtargeted to nucleotides 22208-22235 of SEQ ID NO: 2 is selected fromISIS NOs: 405992, 405993, 405994, 405995, 410556, 410557, 410558,410559, 410560, 410623, 410624, 410625, 410626, 410627, 410628, 410629,410630, 410631, 410703, 410704, 410705, 410706, 410707, 410708, 410709,410710 or 410711.

In certain embodiments, a target region is nucleotides 22209-22236 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22209-22236 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401,402, 403, 404 or 405. In certain such embodiments, an antisense compoundtargeted to nucleotides 22209-22236 of SEQ ID NO: 2 is selected fromISIS NOs: 405993, 405994, 405995, 410556, 410557, 410558, 410559,410560, 410561, 410624, 410625, 410626, 410627, 410628, 410629, 410630,410631, 410632, 410704, 410705, 410706, 410707, 410708, 410709, 410710,410711 or 410712.

In certain embodiments, a target region is nucleotides 22210-22236 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22210-22236 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402,403, 404 or 405. In certain such embodiments, an antisense compoundtargeted to nucleotides 22210-22236 of SEQ ID NO: 2 is selected fromISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560,410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632,410705, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.

In certain embodiments, a target region is nucleotides 22210-22239 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22210-22239 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402,403, 404, 405 or 405. In certain such embodiments, an antisense compoundtargeted to nucleotides 22210-22239 of SEQ ID NO: 2 is selected fromISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560,410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632,410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712 or410774.

In certain embodiments, a target region is nucleotides 22211-22236 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22211-22236 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403,404 or 405. In certain such embodiments, an antisense compound targetedto nucleotides 22211-22236 of SEQ ID NO: 2 is selected from ISIS NOs:405994, 405995, 410557, 410558, 410559, 410560, 410561, 410626, 410627,410628, 410629, 410630, 410631, 410632, 410706, 410707, 410708, 410709,410710, 410711 or 410712.

In certain embodiments, a target region is nucleotides 22212-22239 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22212-22239 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404,405 or 406. In certain such embodiments, an antisense compound targetedto nucleotides 22212-22239 of SEQ ID NO: 2 is selected from ISIS NOs:405994, 405995, 410558, 410559, 410560, 410561, 410627, 410628, 410629,410630, 410631, 410632, 410707, 410708, 410709, 410710, 410711, 410712or 410774.

In certain embodiments, a target region is nucleotides 22292-22311 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 22292-22311 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 126. In certain suchembodiments, an antisense compound targeted to nucleotides 22292-22311of SEQ ID NO: 2 is selected from ISIS NOs: 395225 or 399947.

In certain embodiments, a target region is nucleotides 23985-24054 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 23985-24054 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 408, 409 or 410. Incertain such embodiments, an antisense compound targeted to nucleotides23985-24054 of SEQ ID NO: 2 is selected from ISIS NOs: 410776, 410777 or410778.

In certain embodiments, a target region is nucleotides 24035-24134 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 24035-24134 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412,413, 414, 415 or 416. In certain such embodiments, an antisense compoundtargeted to nucleotides 24035-24134 of SEQ ID NO: 2 is selected fromISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562,410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715,410716, 410717, 410718, 410778 or 410779.

In certain embodiments, a target region is nucleotides 24095-24121 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 24095-24121 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414or 415. In certain such embodiments, an antisense compound targeted tonucleotides 24095-24121 of SEQ ID NO: 2 is selected from ISIS NOs:395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564,410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717or 410718.

In certain embodiments, a target region is nucleotides 24858-24877 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 24858-24877 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 147. In certain suchembodiments, an antisense compound targeted to nucleotides 24858-24877of SEQ ID NO: 2 is selected from ISIS NOs: 395226 or 399948.

In certain embodiments, a target region is nucleotides 24907-24926 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 24907-24926 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 118. In certain suchembodiments, an antisense compound targeted to nucleotides 24907-24926of SEQ ID NO: 2 is selected from ISIS NOs: 399869 or 400025.

In certain embodiments, a target region is nucleotides 25413-25432 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 25413-25432 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 114. In certain suchembodiments, an antisense compound targeted to nucleotides 25413-25432of SEQ ID NO: 2 is selected from ISIS NOs: 399870 or 400026.

In certain embodiments, a target region is nucleotides 25994-26013 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 25994-26013 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 418. In certain suchembodiments, an antisense compound targeted to nucleotides 25994-26013of SEQ ID NO: 2 is selected from ISIS NOs: 410781.

In certain embodiments, a target region is nucleotides 26112-26139 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26112-26139 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425 or 426. In certain such embodiments, an antisense compoundtargeted to nucleotides 26112-26139 of SEQ ID NO: 2 is selected fromISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568,410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642,410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725,410726 or 410727.

In certain embodiments, a target region is nucleotides 26112-26161 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26112-26161 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425, 426, 427, 428, 429 or 430. In certain such embodiments,an antisense compound targeted to nucleotides 26112-26161 of SEQ ID NO:2 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566,410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638,410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719,410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728,410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26112-27303 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26112-27303 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 112, 122, 135, 153, 154, 419,420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433,434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445 or 446. Incertain such embodiments, an antisense compound targeted to nucleotides26112-27303 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188,395189, 395190, 395191, 395192, 395193, 395194, 395195, 395196, 395197,395198, 395199, 399818, 399819, 399820, 399821, 399822, 399823, 399824,399825, 399909, 399910, 399911, 399912, 399913, 399914, 399915, 399916,399917, 399918, 399919, 399920, 399921, 399974, 399975, 399976, 399977,399978, 399979, 399980, 399981, 405893, 405894, 405895, 405896, 405897,405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906,405907, 405908, 405998, 410565, 410566, 410567, 410568, 410569, 410570,410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642,410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723,410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26113-26140 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26113-26140 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423,424, 425, 426 or 427. In certain such embodiments, an antisense compoundtargeted to nucleotides 26113-26140 of SEQ ID NO: 2 is selected fromISIS NOs: 395187, 399909, 405998, 410566, 410567, 410568, 410569,410570, 410571, 410572, 410638, 410639, 410640, 410641, 410642, 410643,410644, 410645, 410720, 410721, 410722, 410723, 410724, 410725, 410726,410727 or 410728.

In certain embodiments, a target region is nucleotides 26114-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26114-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424,425, 426, 427 or 248. In certain such embodiments, an antisense compoundtargeted to nucleotides 26114-26141 of SEQ ID NO: 2 is selected fromISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570,410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644,410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727,410728 or 410729.

In certain embodiments, a target region is nucleotides 26115-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26115-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425,426, 427 or 248. In certain such embodiments, an antisense compoundtargeted to nucleotides 26115-26141 of SEQ ID NO: 2 is selected fromISIS NOs: 395187, 399909, 405998, 410568, 410569, 410570, 410571,410572, 410573, 410640, 410641, 410642, 410643, 410644, 410645, 410646,410722, 410723, 410724, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26116-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26116-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426,427 or 428. In certain such embodiments, an antisense compound targetedto nucleotides 26116-26141 of SEQ ID NO: 2 is selected from ISIS NOs:395187, 399909, 405998, 410569, 410570, 410571, 410572, 410573, 410641,410642, 410643, 410644, 410645, 410646, 410723, 410724, 410725, 410726,410727 or 410728.

In certain embodiments, a target region is nucleotides 26117-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26117-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427 or428. In certain such embodiments, an antisense compound targeted tonucleotides 26117-26141 of SEQ ID NO: 2 is selected from ISIS NOs:395187, 399909, 405998, 410570, 410571, 410572, 410573, 410642, 410643,410644, 410645, 410646, 410724, 410725, 410726, 410727, 410728 or410729.

In certain embodiments, a target region is nucleotides 26117-26475 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26117-26475 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67,122, 153, 154, 424, 425, 426, 427, 428, 429 or 430. In certain suchembodiments, an antisense compound targeted to nucleotides 26117-26475of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188, 395189,395190, 395191, 395192, 399818, 399819, 399820, 399909, 399910, 399911,399912, 399913, 399914, 399974, 399975, 399976, 405998, 410570, 410571,410572, 410573, 410642, 410643, 410644, 410645, 410646, 410724, 410725,410726, 410727, 410728, 410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26118-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26118-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427 or 428.In certain such embodiments, an antisense compound targeted tonucleotides 26118-26141 of SEQ ID NO: 2 is selected from ISIS NOs:405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645,410646, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26120-26141 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26120-26141 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. Incertain such embodiments, an antisense compound targeted to nucleotides26120-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 405998, 410572,410573, 410644, 410645, 410646, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26132-26151 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26132-26151 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 429. In certain suchembodiments, an antisense compound targeted to nucleotides 26132-26151of SEQ ID NO: 2 is selected from ISIS NOs: 410782.

In certain embodiments, a target region is nucleotides 26142-26161 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26142-26161 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 430. In certain suchembodiments, an antisense compound targeted to nucleotides 26142-26161of SEQ ID NO: 2 is selected from ISIS NOs: 410783.

In certain embodiments, a target region is nucleotides 26217-26241 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26217-26241 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain suchembodiments, an antisense compound targeted to nucleotides 26217-26241of SEQ ID NO: 2 is selected from ISIS NOs: 395188, 399818 or 399910.

In certain embodiments, a target region is nucleotides 26311-26335 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26311-26335 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain suchembodiments, an antisense compound targeted to nucleotides 26311-26335of SEQ ID NO: 2 is selected from ISIS NOs: 395189, 399819, 399911 or399975.

In certain embodiments, a target region is nucleotides 26389-26432 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26389-26432 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certainsuch embodiments, an antisense compound targeted to nucleotides26389-26432 of SEQ ID NO: 2 is selected from ISIS NOs: 395190, 395191,399820, 399912, 399913 or 399976.

In certain embodiments, a target region is nucleotides 26456-26576 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26456-26576 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 68 or 153. In certain suchembodiments, an antisense compound targeted to nucleotides 26456-26576of SEQ ID NO: 2 is selected from ISIS NOs: 395192, 395193, 399914 or399915.

In certain embodiments, a target region is nucleotides 26635-26662 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26635-26662 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434,435, 436, 437 or 438. In certain such embodiments, an antisense compoundtargeted to nucleotides 26635-26662 of SEQ ID NO: 2 is selected fromISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897,405898, 405899 or 405900.

In certain embodiments, a target region is nucleotides 26707-26734 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26707-26734 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441,442, 443 or 444. In certain such embodiments, an antisense compoundtargeted to nucleotides 26707-26734 of SEQ ID NO: 2 is selected fromISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903,405904, 405905 or 405906.

In certain embodiments, a target region is nucleotides 26707-26736 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26707-26736 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441,442, 443, 444, 445 or 446. In certain such embodiments, an antisensecompound targeted to nucleotides 26707-26736 of SEQ ID NO: 2 is selectedfrom ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903,405904, 405905, 405906, 405907 or 405908.

In certain embodiments, a target region is nucleotides 26790-26820 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 26790-26820 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certainsuch embodiments, an antisense compound targeted to nucleotides26790-26820 of SEQ ID NO: 2 is selected from ISIS NOs: 395196, 399822,399823, 399918, 399978 or 399979.

In certain embodiments, a target region is nucleotides 27034-27263 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 27034-27263 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certainsuch embodiments, an antisense compound targeted to nucleotides27034-27263 of SEQ ID NO: 2 is selected from ISIS NOs: 395197, 395198,399824, 399919, 399920 or 399980.

In certain embodiments, a target region is nucleotides 27279-27303 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 27279-27303 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain suchembodiments, an antisense compound targeted to nucleotides 27279-27303of SEQ ID NO: 2 is selected from ISIS NOs: 395199, 399825, 399921 or399981.

In certain embodiments, a target region is nucleotides 27350-27376 ofSEQ ID NO: 2. In certain embodiments, an antisense compound is targetedto nucleotides 27350-27376 of SEQ ID NO: 2. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain suchembodiments, an antisense compound targeted to nucleotides 27350-27376of SEQ ID NO: 2 is selected from ISIS NOs: 395200, 399826, 399922 or399982.

In certain embodiments, antisense compounds target a PCSK9 nucleic acidhaving the sequence of GENBANK® Accession No. AK124635.1, firstdeposited with GENBANK® on Sep. 8, 2003, and incorporated herein as SEQID NO: 3. In certain such embodiments, an antisense oligonucleotide istargeted to SEQ ID NO: 3. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 3 is at least 90%complementary to SEQ ID NO: 1. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 3 is at least 95%complementary to SEQ ID NO: 3. In certain such embodiments, an antisenseoligonucleotide that is targeted to SEQ ID NO: 3 is 100% complementaryto SEQ ID NO: 3. In certain such embodiments, an antisenseoligonucleotide comprises a nucleotide sequence selected from anucleotide sequence set forth in Table 12.

TABLE 12 Nucleotide sequences targeted to AK124635.1 (SEQ ID NO: 3)5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID NO NO: 3 NO: 3Sequences (5′-3′) 85 155 174 ACAAATTCCCAGACTCAGCA 100 220 239ATCTCAGGACAGGTGAGCAA 116 234 253 GAGTAGAGATTCTCATCTCA 129 290 309GTGCCATCTGAACAGCACCT 117 302 321 GAGTCTTCTGAAGTGCCATC 81 377 396AAGCAGGGCCTCAGGTGGAA 110 400 419 CCTGGAACCCCTGCAGCCAG 152 451 470TTCAGGCAGGTTGCTGCTAG 83 460 479 AAGGAAGACTTCAGGCAGGT 140 472 491TCAGCCAGGCCAAAGGAAGA 137 586 605 TAGGGAGAGCTCACAGATGC 136 596 615TAGGAGAAAGTAGGGAGAGC 132 653 672 TAAAAGCTGCAAGAGACTCA 139 741 760TCAGAGAAAACAGTCACCGA 92 871 890 AGAGACAGGAAGCTGCAGCT 142 878 897TCATTTTAGAGACAGGAAGC 113 915 934 GAATAACAGTGATGTCTGGC 138 968 987TCACAGCTCACCGAGTCTGC 98 998 1017 AGTGTAAAATAAAGCCCCTA 96 1075 1094AGGACCCAAGTCATCCTGCT 124 1105 1124 GGCCATCAGCTGGCAATGCT 82 1144 1163AAGGAAAGGGAGGCCTAGAG 133 1149 1168 TAGACAAGGAAAGGGAGGCC 103 1155 1174ATTTCATAGACAAGGAAAGG 155 1275 1294 CTTATAGTTAACACACAGAA 156 1283 1302AAGTCAACCTTATAGTTAAC 146 1315 1334 TGACATTTGTGGGAGAGGAG 161 1322 1341TCCAAGGTGACATTTGTGGG 215 1351 1370 ATACACCTCCACCAGGCTGC 216 1362 1381GTGTCTAGGAGATACACCTC 19 1365 1384 CTGGTGTCTAGGAGATACAC 217 1367 1386TGCTGGTGTCTAGGAGATAC 20 1370 1389 GTATGCTGGTGTCTAGGAGA 21 1390 1409GATTTCCCGGTGGTCACTCT 218 1392 1411 TCGATTTCCCGGTGGTCACT 219 1393 1412CTCGATTTCCCGGTGGTCAC 220 1394 1413 CCTCGATTTCCCGGTGGTCA 221 1395 1414CCCTCGATTTCCCGGTGGTC 22 1396 1415 GCCCTCGATTTCCCGGTGGT 222 1397 1416TGCCCTCGATTTCCCGGTGG 223 1398 1417 CTGCCCTCGATTTCCCGGTG 224 1399 1418CCTGCCCTCGATTTCCCGGT 225 1400 1419 CCCTGCCCTCGATTTCCCGG 226 1404 1423ATGACCCTGCCCTCGATTTC 227 1406 1425 CCATGACCCTGCCCTCGATT 228 1408 1427GACCATGACCCTGCCCTCGA 23 1410 1429 GTGACCATGACCCTGCCCTC 229 1412 1431CGGTGACCATGACCCTGCCC 230 1414 1433 GTCGGTGACCATGACCCTGC 231 1416 1435AAGTCGGTGACCATGACCCT 232 1418 1437 CGAAGTCGGTGACCATGACC 24 1420 1439CTCGAAGTCGGTGACCATGA 233 1428 1447 GGCACATTCTCGAAGTCGGT 25 1453 1472GTGGAAGCGGGTCCCGTCCT 234 1463 1482 TGGCCTGTCTGTGGAAGCGG 235 1490 1509GGTGGGTGCCATGACTGTCA 236 1493 1512 CCAGGTGGGTGCCATGACTG 237 1497 1516CCTGCCAGGTGGGTGCCATG 26 1500 1519 ACCCCTGCCAGGTGGGTGCC 238 1502 1521CCACCCCTGCCAGGTGGGTG 27 1505 1524 TGACCACCCCTGCCAGGTGG 239 1507 1526GCTGACCACCCCTGCCAGGT 240 1515 1534 TCCCGGCCGCTGACCACCCC 241 1519 1538GGCATCCCGGCCGCTGACCA 242 1522 1541 GCCGGCATCCCGGCCGCTGA 243 1527 1546GCCACGCCGGCATCCCGGCC 244 1528 1547 GGCCACGCCGGCATCCCGGC 245 1529 1548TGGCCACGCCGGCATCCCGG 246 1530 1549 TTGGCCACGCCGGCATCCCG 247 1531 1550CTTGGCCACGCCGGCATCCC 248 1532 1551 CCTTGGCCACGCCGGCATCC 249 1533 1552CCCTTGGCCACGCCGGCATC 28 1534 1553 ACCCTTGGCCACGCCGGCAT 447 1534 1553ACCCTTGGTCACGCCGGCAT 250 1535 1554 CACCCTTGGCCACGCCGGCA 251 1536 1555GCACCCTTGGCCACGCCGGC 252 1537 1556 GGCACCCTTGGCCACGCCGG 253 1538 1557TGGCACCCTTGGCCACGCCG 254 1539 1558 CTGGCACCCTTGGCCACGCC 255 1540 1559GCTGGCACCCTTGGCCACGC 256 1545 1564 CGCATGCTGGCACCCTTGGC 257 1566 1585CAGTTGAGCACGCGCAGGCT 258 1568 1587 GGCAGTTGAGCACGCGCAGG 29 1570 1589TTGGCAGTTGAGCACGCGCA 259 1572 1591 CCTTGGCAGTTGAGCACGCG 30 1575 1594TTCCCTTGGCAGTTGAGCAC 260 1577 1596 CCTTCCCTTGGCAGTTGAGC 261 1581 1600GTGCCCTTCCCTTGGCAGTT 262 1583 1602 CCGTGCCCTTCCCTTGGCAG 263 1594 1613GGTGCCGCTAACCGTGCCCT 264 1606 1625 CAGGCCTATGAGGGTGCCGC 31 1607 1626CCAGGCCTATGAGGGTGCCG 458 1609 1622 GCCTATGAGGGTGC 459 1614 1627TCCAGGCCTATGAG 265 1618 1637 CCGAATAAACTCCAGGCCTA 266 1626 1645TGGCTTTTCCGAATAAACTC 32 1628 1647 GCTGGCTTTTCCGAATAAAC 267 1630 1649CAGCTGGCTTTTCCGAATAA 268 1632 1651 ACCAGCTGGCTTTTCCGAAT 269 1634 1653GGACCAGCTGGCTTTTCCGA 270 1638 1657 GGCTGGACCAGCTGGCTTTT 271 1649 1668GTGGCCCCACAGGCTGGACC 272 1662 1681 AGCAGCACCACCAGTGGCCC 273 1684 1703GCTGTACCCACCCGCCAGGG 274 1730 1749 CGACCCCAGCCCTCGCCAGG 33 1740 1759GTGACCAGCACGACCCCAGC 275 1742 1761 CGGTGACCAGCACGACCCCA 276 1744 1763AGCGGTGACCAGCACGACCC 277 1746 1765 GCAGCGGTGACCAGCACGAC 278 1748 1767CGGCAGCGGTGACCAGCACG 279 1749 1768 CCGGCAGCGGTGACCAGCAC 280 1752 1771TTGCCGGCAGCGGTGACCAG 281 1754 1773 AGTTGCCGGCAGCGGTGACC 282 1756 1775GAAGTTGCCGGCAGCGGTGA 283 1758 1777 CGGAAGTTGCCGGCAGCGGT 284 1760 1779CCCGGAAGTTGCCGGCAGCG 285 1762 1781 GTCCCGGAAGTTGCCGGCAG 306 1820 1839GAGGCACCAATGATGTCCTC 39 1822 1841 TGGAGGCACCAATGATGTCC 307 1824 1843GCTGGAGGCACCAATGATGT 308 1826 1845 TCGCTGGAGGCACCAATGAT 309 1828 1847AGTCGCTGGAGGCACCAATG 310 1830 1849 GCAGTCGCTGGAGGCACCAA 40 1833 1852GCTGCAGTCGCTGGAGGCAC 311 1835 1854 GTGCTGCAGTCGCTGGAGGC 312 1837 1856AGGTGCTGCAGTCGCTGGAG 313 1839 1858 GCAGGTGCTGCAGTCGCTGG 314 1840 1859AGCAGGTGCTGCAGTCGCTG 315 1842 1861 AAAGCAGGTGCTGCAGTCGC 41 1844 1863ACAAAGCAGGTGCTGCAGTC 316 1846 1865 ACACAAAGCAGGTGCTGCAG 317 1848 1867TGACACAAAGCAGGTGCTGC 318 1858 1877 TCCCACTCTGTGACACAAAG 101 1898 1917ATGGCTGCAATGCCAGCCAC 319 1900 1919 TCATGGCTGCAATGCCAGCC 42 1903 1922GCATCATGGCTGCAATGCCA 320 1905 1924 CAGCATCATGGCTGCAATGC 321 1907 1926GACAGCATCATGGCTGCAAT 322 1909 1928 CAGACAGCATCATGGCTGCA 43 1911 1930GGCAGACAGCATCATGGCTG 323 1913 1932 TCGGCAGACAGCATCATGGC 324 1915 1934GCTCGGCAGACAGCATCATG 325 1917 1936 CGGCTCGGCAGACAGCATCA 326 1919 1938TCCGGCTCGGCAGACAGCAT 327 1933 1952 CGGCCAGGGTGAGCTCCGGC 328 1946 1965CTCTGCCTCAACTCGGCCAG 329 1948 1967 GTCTCTGCCTCAACTCGGCC 330 1950 1969CAGTCTCTGCCTCAACTCGG 331 1952 1971 ATCAGTCTCTGCCTCAACTC 332 1954 1973GGATCAGTCTCTGCCTCAAC 333 1956 1975 GTGGATCAGTCTCTGCCTCA 334 1958 1977AAGTGGATCAGTCTCTGCCT 44 1959 1978 GAAGTGGATCAGTCTCTGCC 335 1961 1980GAGAAGTGGATCAGTCTCTG 336 1963 1982 CAGAGAAGTGGATCAGTCTC 337 1965 1984GGCAGAGAAGTGGATCAGTC 45 1967 1986 TTGGCAGAGAAGTGGATCAG 338 1969 1988CTTTGGCAGAGAAGTGGATC 46 1972 1991 CATCTTTGGCAGAGAAGTGG 339 1974 1993GACATCTTTGGCAGAGAAGT 340 1976 1995 ATGACATCTTTGGCAGAGAA 47 1978 1997TGATGACATCTTTGGCAGAG 341 1980 1999 ATTGATGACATCTTTGGCAG 342 1982 2001TCATTGATGACATCTTTGGC 48 1985 2004 GCCTCATTGATGACATCTTT 343 1987 2006AGGCCTCATTGATGACATCT 344 1989 2008 CCAGGCCTCATTGATGACAT 345 1991 2010AACCAGGCCTCATTGATGAC 346 1993 2012 GGAACCAGGCCTCATTGATG 347 1994 2013GGGAACCAGGCCTCATTGAT 348 1995 2014 AGGGAACCAGGCCTCATTGA 349 1996 2015CAGGGAACCAGGCCTCATTG 49 1997 2016 TCAGGGAACCAGGCCTCATT 350 1998 2017CTCAGGGAACCAGGCCTCAT 351 1999 2018 CCTCAGGGAACCAGGCCTCA 352 2000 2019TCCTCAGGGAACCAGGCCTC 353 2001 2020 GTCCTCAGGGAACCAGGCCT 50 2002 2021GGTCCTCAGGGAACCAGGCC 354 2003 2022 TGGTCCTCAGGGAACCAGGC 355 2004 2023CTGGTCCTCAGGGAACCAGG 356 2005 2024 GCTGGTCCTCAGGGAACCAG 357 2006 2025CGCTGGTCCTCAGGGAACCA 87 2009 2028 ACCCGCTGGTCCTCAGGGAA 358 2011 2030GTACCCGCTGGTCCTCAGGG 359 2013 2032 CAGTACCCGCTGGTCCTCAG 51 2016 2035GGTCAGTACCCGCTGGTCCT 119 2038 2057 GCAGGGCGGCCACCAGGTTG 360 2061 2080ACCTGCCCCATGGGTGCTGG 52 2073 2092 AAACAGCTGCCAACCTGCCC 361 2075 2094CAAAACAGCTGCCAACCTGC 53 2078 2097 CTGCAAAACAGCTGCCAACC 362 2080 2099TCCTGCAAAACAGCTGCCAA 363 2082 2101 AGTCCTGCAAAACAGCTGCC 104 2085 2104CACAGTCCTGCAAAACAGCT 130 2095 2114 GTGCTGACCACACAGTCCTG 365 2105 2124GGCCCCGAGTGTGCTGACCA 54 2108 2127 GTAGGCCCCGAGTGTGCTGA 366 2110 2129GTGTAGGCCCCGAGTGTGCT 367 2112 2131 CCGTGTAGGCCCCGAGTGTG 368 2114 2133ATCCGTGTAGGCCCCGAGTG 369 2116 2135 CCATCCGTGTAGGCCCCGAG 370 2118 2137GGCCATCCGTGTAGGCCCCG 371 2120 2139 GTGGCCATCCGTGTAGGCCC 372 2168 2187CTGGAGCAGCTCAGCAGCTC 460 2168 2181 CAGCTCAGCAGCTC 373 2170 2189AACTGGAGCAGCTCAGCAGC 55 2173 2192 AGAAACTGGAGCAGCTCAGC 374 2175 2194GGAGAAACTGGAGCAGCTCA 375 2177 2196 CTGGAGAAACTGGAGCAGCT 56 2179 2198TCCTGGAGAAACTGGAGCAG 408 2245 2264 GTGCCAAGGTCCTCCACCTC 409 2265 2284TCAGCACAGGCGGCTTGTGG 410 2295 2314 CCACGCACTGGTTGGGCTGA 60 2355 2374CTTTGCATTCCAGACCTGGG 411 2356 2375 ACTTTGCATTCCAGACCTGG 412 2357 2376GACTTTGCATTCCAGACCTG 413 2358 2377 TGACTTTGCATTCCAGACCT 414 2359 2378TTGACTTTGCATTCCAGACC 61 2360 2379 CTTGACTTTGCATTCCAGAC 415 2362 2381TCCTTGACTTTGCATTCCAG 416 2375 2394 CGGGATTCCATGCTCCTTGA 417 2405 2424GCAGGCCACGGTCACCTGCT 418 2442 2461 GGAGGGCACTGCAGCCAGTC 419 2560 2579CAGATGGCAACGGCTGTCAC 420 2561 2580 GCAGATGGCAACGGCTGTCA 421 2562 2581AGCAGATGGCAACGGCTGTC 422 2563 2582 CAGCAGATGGCAACGGCTGT 423 2564 2583GCAGCAGATGGCAACGGCTG 62 2565 2584 GGCAGCAGATGGCAACGGCT 424 2566 2585CGGCAGCAGATGGCAACGGC 425 2567 2586 CCGGCAGCAGATGGCAACGG 426 2568 2587TCCGGCAGCAGATGGCAACG 427 2569 2588 CTCCGGCAGCAGATGGCAAC 428 2570 2589GCTCCGGCAGCAGATGGCAA 429 2580 2599 CCAGGTGCCGGCTCCGGCAG 430 2590 2609GAGGCCTGCGCCAGGTGCCG 154 2665 2684 TTTTAAAGCTCAGCCCCAGC 63 2670 2689AACCATTTTAAAGCTCAGCC 64 2759 2778 TCAAGGGCCAGGCCAGCAGC 65 2764 2783CCCACTCAAGGGCCAGGCCA 122 2837 2856 GGAGGGAGCTTCCTGGCACC 66 2852 2871ATGCCCCACAGTGAGGGAGG 67 2861 2880 AATGGTGAAATGCCCCACAG 153 2904 2923TTGGGAGCAGCTGGCAGCAC 68 3005 3024 CATGGGAAGAATCCTGCCTC 431 3083 3102ATGAGGGCCATCAGCACCTT 432 3084 3103 GATGAGGGCCATCAGCACCT 433 3085 3104AGATGAGGGCCATCAGCACC 434 3086 3105 GAGATGAGGGCCATCAGCAC 69 3087 3106GGAGATGAGGGCCATCAGCA 435 3088 3107 TGGAGATGAGGGCCATCAGC 436 3089 3108CTGGAGATGAGGGCCATCAG 437 3090 3109 GCTGGAGATGAGGGCCATCA 438 3091 3110AGCTGGAGATGAGGGCCATC 461 3132 3145 TTAATCAGGGAGCC 70 3155 3174TAGATGCCATCCAGAAAGCT 439 3157 3176 GCTAGATGCCATCCAGAAAG 440 3158 3177GGCTAGATGCCATCCAGAAA 441 3159 3178 TGGCTAGATGCCATCCAGAA 442 3160 3179CTGGCTAGATGCCATCCAGA 71 3161 3180 TCTGGCTAGATGCCATCCAG 443 3162 3181CTCTGGCTAGATGCCATCCA 444 3163 3182 CCTCTGGCTAGATGCCATCC 445 3164 3183GCCTCTGGCTAGATGCCATC 446 3165 3184 AGCCTCTGGCTAGATGCCAT 72 3238 3257GGCATAGAGCAGAGTAAAGG 73 3243 3262 AGCCTGGCATAGAGCAGAGT 135 3249 3268TAGCACAGCCTGGCATAGAG 112 3482 3501 GAAGAGGCTTGGCTTCAGAG 74 3488 3507AAGTAAGAAGAGGCTTGGCT 75 3692 3711 GCTCAAGGAGGGACAGTTGT 76 3727 3746AAAGATAAATGTCTGCTTGC 77 3732 3751 ACCCAAAAGATAAATGTCTG 78 3798 3817TCTTCAAGTTACAAAAGCAA 99 3805 3824 ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to aPCSK9 nucleic acid. In certain such embodiments, gapmer antisensecompounds are targeted to SEQ ID NO: 3. In certain such embodiments, thenucleotide sequences illustrated in Table 12 have a 5-10-5 gapmer motifTable 13 illustrates gapmer antisense compounds targeted to SEQ ID NO:3, having a 5-10-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 13 Gapmer antisense compounds having a 5-10-5 motif targeted toSEQ ID NO: 3 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ IDIsis No Motif NO NO: 3 NO: 3 Mismatches Sequence (5′-3′) 395201 5-10-585 155 174 0 ACAAATTCCCAGACTCAGCA 399827 5-10-5 100 220 239 0ATCTCAGGACAGGTGAGCAA 399828 5-10-5 116 234 253 0 GAGTAGAGATTCTCATCTCA395202 5-10-5 129 290 309 0 GTGCCATCTGAACAGCACCT 399829 5-10-5 117 302321 0 GAGTCTTCTGAAGTGCCATC 399830 5-10-5 81 377 396 0AAGCAGGGCCTCAGGTGGAA 395203 5-10-5 110 400 419 0 CCTGGAACCCCTGCAGCCAG395204 5-10-5 152 451 470 0 TTCAGGCAGGTTGCTGCTAG 399831 5-10-5 83 460479 0 AAGGAAGACTTCAGGCAGGT 395205 5-10-5 140 472 491 0TCAGCCAGGCCAAAGGAAGA 399832 5-10-5 137 586 605 0 TAGGGAGAGCTCACAGATGC395206 5-10-5 136 596 615 0 TAGGAGAAAGTAGGGAGAGC 395207 5-10-5 132 653672 0 TAAAAGCTGCAAGAGACTCA 395208 5-10-5 139 741 760 0TCAGAGAAAACAGTCACCGA 399833 5-10-5 92 871 890 0 AGAGACAGGAAGCTGCAGCT395209 5-10-5 142 878 897 0 TCATTTTAGAGACAGGAAGC 395210 5-10-5 113 915934 0 GAATAACAGTGATGTCTGGC 395211 5-10-5 138 968 987 0TCACAGCTCACCGAGTCTGC 395212 5-10-5 98 998 1017 0 AGTGTAAAATAAAGCCCCTA395213 5-10-5 96 1075 1094 0 AGGACCCAAGTCATCCTGCT 395214 5-10-5 124 11051124 0 GGCCATCAGCTGGCAATGCT 399834 5-10-5 82 1144 1163 0AAGGAAAGGGAGGCCTAGAG 395215 5-10-5 133 1149 1168 0 TAGACAAGGAAAGGGAGGCC395216 5-10-5 103 1155 1174 0 ATTTCATAGACAAGGAAAGG 395217 5-10-5 1551275 1294 0 CTTATAGTTAACACACAGAA 399835 5-10-5 156 1283 1302 0AAGTCAACCTTATAGTTAAC 395218 5-10-5 146 1315 1334 0 TGACATTTGTGGGAGAGGAG395219 5-10-5 161 1322 1341 0 TCCAAGGTGACATTTGTGGG 395160 5-10-5 19 13651384 0 CTGGTGTCTAGGAGATACAC 399797 5-10-5 20 1370 1389 0GTATGCTGGTGTCTAGGAGA 395161 5-10-5 21 1390 1409 0 GATTTCCCGGTGGTCACTCT399798 5-10-5 22 1396 1415 0 GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 14101429 0 GTGACCATGACCCTGCCCTC 395162 5-10-5 24 1420 1439 0CTCGAAGTCGGTGACCATGA 395163 5-10-5 25 1453 1472 0 GTGGAAGCGGGTCCCGTCCT395164 5-10-5 26 1500 1519 0 ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 15051524 0 TGACCACCCCTGCCAGGTGG 395165 5-10-5 28 1534 1553 0ACCCTTGGCCACGCCGGCAT 395166 5-10-5 29 1570 1589 0 TTGGCAGTTGAGCACGCGCA399801 5-10-5 30 1575 1594 0 TTCCCTTGGCAGTTGAGCAC 395167 5-10-5 31 16071626 0 CCAGGCCTATGAGGGTGCCG 395168 5-10-5 32 1628 1647 0GCTGGCTTTTCCGAATAAAC 395169 5-10-5 33 1740 1759 0 GTGACCAGCACGACCCCAGC395175 5-10-5 39 1822 1841 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 18331852 0 GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 1844 1863 0ACAAAGCAGGTGCTGCAGTC 395176 5-10-5 101 1898 1917 0 ATGGCTGCAATGCCAGCCAC399806 5-10-5 42 1903 1922 0 GCATCATGGCTGCAATGCCA 399807 5-10-5 43 19111930 0 GGCAGACAGCATCATGGCTG 399808 5-10-5 44 1959 1978 0GAAGTGGATCAGTCTCTGCC 395177 5-10-5 45 1967 1986 0 TTGGCAGAGAAGTGGATCAG399809 5-10-5 46 1972 1991 0 CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 19781997 0 TGATGACATCTTTGGCAGAG 399811 5-10-5 48 1985 2004 0GCCTCATTGATGACATCTTT 399812 5-10-5 49 1997 2016 0 TCAGGGAACCAGGCCTCATT395178 5-10-5 50 2002 2021 0 GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 20092028 0 ACCCGCTGGTCCTCAGGGAA 399813 5-10-5 51 2016 2035 0GGTCAGTACCCGCTGGTCCT 395180 5-10-5 119 2038 2057 0 GCAGGGCGGCCACCAGGTTG395181 5-10-5 52 2073 2092 0 AAACAGCTGCCAACCTGCCC 399814 5-10-5 53 20782097 0 CTGCAAAACAGCTGCCAACC 395220 5-10-5 104 2085 2104 0CACAGTCCTGCAAAACAGCT 399836 5-10-5 130 2095 2114 0 GTGCTGACCACACAGTCCTG395182 5-10-5 54 2108 2127 0 GTAGGCCCCGAGTGTGCTGA 399815 5-10-5 55 21732192 0 AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 2179 2198 0TCCTGGAGAAACTGGAGCAG 395186 5-10-5 60 2355 2374 0 CTTTGCATTCCAGACCTGGG399817 5-10-5 61 2360 2379 0 CTTGACTTTGCATTCCAGAC 395187 5-10-5 62 25652584 0 GGCAGCAGATGGCAACGGCT 395188 5-10-5 154 2665 2684 0TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 63 2670 2689 0 AACCATTTTAAAGCTCAGCC395189 5-10-5 64 2759 2778 0 TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 27642783 0 CCCACTCAAGGGCCAGGCCA 399820 5-10-5 122 2837 2856 0GGAGGGAGCTTCCTGGCACC 395190 5-10-5 66 2852 2871 0 ATGCCCCACAGTGAGGGAGG395191 5-10-5 67 2861 2880 0 AATGGTGAAATGCCCCACAG 395192 5-10-5 153 29042923 0 TTGGGAGCAGCTGGCAGCAC 395193 5-10-5 68 3005 3024 0CATGGGAAGAATCCTGCCTC 395194 5-10-5 69 3087 3106 0 GGAGATGAGGGCCATCAGCA395195 5-10-5 70 3155 3174 0 TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 31613180 0 TCTGGCTAGATGCCATCCAG 395196 5-10-5 72 3238 3257 0GGCATAGAGCAGAGTAAAGG 399822 5-10-5 73 3243 3262 0 AGCCTGGCATAGAGCAGAGT399823 5-10-5 135 3249 3268 0 TAGCACAGCCTGGCATAGAG 395197 5-10-5 1123482 3501 0 GAAGAGGCTTGGCTTCAGAG 399824 5-10-5 74 3488 3507 0AAGTAAGAAGAGGCTTGGCT 395198 5-10-5 75 3692 3711 0 GCTCAAGGAGGGACAGTTGT395199 5-10-5 76 3727 3746 0 AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 37323751 0 ACCCAAAAGATAAATGTCTG 395200 5-10-5 78 3798 3817 0TCTTCAAGTTACAAAAGCAA 399826 5-10-5 99 3805 3824 0 ATAAATATCTTCAAGTTACA405869 5-10-5 218 1392 1411 0 TCGATTTCCCGGTGGTCACT 405870 5-10-5 2191393 1412 0 CTCGATTTCCCGGTGGTCAC 405871 5-10-5 220 1394 1413 0CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 1395 1414 0 CCCTCGATTTCCCGGTGGTC405873 5-10-5 222 1397 1416 0 TGCCCTCGATTTCCCGGTGG 405874 5-10-5 2231398 1417 0 CTGCCCTCGATTTCCCGGTG 405875 5-10-5 224 1399 1418 0CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 1400 1419 0 CCCTGCCCTCGATTTCCCGG405877 5-10-5 246 1530 1549 0 TTGGCCACGCCGGCATCCCG 405878 5-10-5 2471531 1550 0 CTTGGCCACGCCGGCATCCC 405879 5-10-5 248 1532 1551 0CCTTGGCCACGCCGGCATCC 405880 5-10-5 249 1533 1552 0 CCCTTGGCCACGCCGGCATC405881 5-10-5 250 1535 1554 0 CACCCTTGGCCACGCCGGCA 405882 5-10-5 2511536 1555 0 GCACCCTTGGCCACGCCGGC 405883 5-10-5 252 1537 1556 0GGCACCCTTGGCCACGCCGG 405884 5-10-5 253 1538 1557 0 TGGCACCCTTGGCCACGCCG405885 5-10-5 346 1993 2012 0 GGAACCAGGCCTCATTGATG 405886 5-10-5 3471994 2013 0 GGGAACCAGGCCTCATTGAT 405887 5-10-5 348 1995 2014 0AGGGAACCAGGCCTCATTGA 405888 5-10-5 349 1996 2015 0 CAGGGAACCAGGCCTCATTG405889 5-10-5 350 1998 2017 0 CTCAGGGAACCAGGCCTCAT 405890 5-10-5 3511999 2018 0 CCTCAGGGAACCAGGCCTCA 405891 5-10-5 352 2000 2019 0TCCTCAGGGAACCAGGCCTC 405892 5-10-5 353 2001 2020 0 GTCCTCAGGGAACCAGGCCT405893 5-10-5 431 3083 3102 0 ATGAGGGCCATCAGCACCTT 405894 5-10-5 4323084 3103 0 GATGAGGGCCATCAGCACCT 405895 5-10-5 433 3085 3104 0AGATGAGGGCCATCAGCACC 405896 5-10-5 434 3086 3105 0 GAGATGAGGGCCATCAGCAC405897 5-10-5 435 3088 3107 0 TGGAGATGAGGGCCATCAGC 405898 5-10-5 4363089 3108 0 CTGGAGATGAGGGCCATCAG 405899 5-10-5 437 3090 3109 0GCTGGAGATGAGGGCCATCA 405900 5-10-5 438 3091 3110 0 AGCTGGAGATGAGGGCCATC405901 5-10-5 439 3157 3176 0 GCTAGATGCCATCCAGAAAG 405902 5-10-5 4403158 3177 0 GGCTAGATGCCATCCAGAAA 405903 5-10-5 441 3159 3178 0TGGCTAGATGCCATCCAGAA 405904 5-10-5 442 3160 3179 0 CTGGCTAGATGCCATCCAGA405905 5-10-5 443 3162 3181 0 CTCTGGCTAGATGCCATCCA 405906 5-10-5 4443163 3182 0 CCTCTGGCTAGATGCCATCC 405907 5-10-5 445 3164 3183 0GCCTCTGGCTAGATGCCATC 405908 5-10-5 446 3165 3184 0 AGCCTCTGGCTAGATGCCAT405909 5-10-5 267 1630 1649 0 CAGCTGGCTTTTCCGAATAA 405910 5-10-5 2681632 1651 0 ACCAGCTGGCTTTTCCGAAT 405911 5-10-5 269 1634 1653 0GGACCAGCTGGCTTTTCCGA 405912 5-10-5 270 1638 1657 0 GGCTGGACCAGCTGGCTTTT405913 5-10-5 275 1742 1761 0 CGGTGACCAGCACGACCCCA 405914 5-10-5 2761744 1763 0 AGCGGTGACCAGCACGACCC 405915 5-10-5 277 1746 1765 0GCAGCGGTGACCAGCACGAC 405916 5-10-5 278 1748 1767 0 CGGCAGCGGTGACCAGCACG405917 5-10-5 280 1752 1771 0 TTGCCGGCAGCGGTGACCAG 405918 5-10-5 2811754 1773 0 AGTTGCCGGCAGCGGTGACC 405919 5-10-5 282 1756 1775 0GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 283 1758 1777 0 CGGAAGTTGCCGGCAGCGGT405921 5-10-5 284 1760 1779 0 CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 2851762 1781 0 GTCCCGGAAGTTGCCGGCAG 405937 5-10-5 306 1820 1839 0GAGGCACCAATGATGTCCTC 405938 5-10-5 307 1824 1843 0 GCTGGAGGCACCAATGATGT405939 5-10-5 308 1826 1845 0 TCGCTGGAGGCACCAATGAT 405940 5-10-5 3091828 1847 0 AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 1830 1849 0GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 1835 1854 0 GTGCTGCAGTCGCTGGAGGC405943 5-10-5 312 1837 1856 0 AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 3141840 1859 0 AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 1842 1861 0AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 1846 1865 0 ACACAAAGCAGGTGCTGCAG405947 5-10-5 317 1848 1867 0 TGACACAAAGCAGGTGCTGC 405948 5-10-5 3191900 1919 0 TCATGGCTGCAATGCCAGCC 405949 5-10-5 320 1905 1924 0CAGCATCATGGCTGCAATGC 405950 5-10-5 321 1907 1926 0 GACAGCATCATGGCTGCAAT405951 5-10-5 322 1909 1928 0 CAGACAGCATCATGGCTGCA 405952 5-10-5 3231913 1932 0 TCGGCAGACAGCATCATGGC 405953 5-10-5 325 1917 1936 0CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 1919 1938 0 TCCGGCTCGGCAGACAGCAT405955 5-10-5 328 1946 1965 0 CTCTGCCTCAACTCGGCCAG 405956 5-10-5 3291948 1967 0 GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 1950 1969 0CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 1952 1971 0 ATCAGTCTCTGCCTCAACTC405959 5-10-5 332 1954 1973 0 GGATCAGTCTCTGCCTCAAC 405960 5-10-5 3331956 1975 0 GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 1958 1977 0AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 1961 1980 0 GAGAAGTGGATCAGTCTCTG405963 5-10-5 337 1965 1984 0 GGCAGAGAAGTGGATCAGTC 405964 5-10-5 3381969 1988 0 CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 1974 1993 0GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 1976 1995 0 ATGACATCTTTGGCAGAGAA405967 5-10-5 341 1980 1999 0 ATTGATGACATCTTTGGCAG 405968 5-10-5 3421982 2001 0 TCATTGATGACATCTTTGGC 405969 5-10-5 343 1987 2006 0AGGCCTCATTGATGACATCT 405970 5-10-5 344 1989 2008 0 CCAGGCCTCATTGATGACAT405971 5-10-5 355 2004 2023 0 CTGGTCCTCAGGGAACCAGG 405972 5-10-5 3572006 2025 0 CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 2011 2030 0GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 2013 2032 0 CAGTACCCGCTGGTCCTCAG405975 5-10-5 361 2075 2094 0 CAAAACAGCTGCCAACCTGC 405976 5-10-5 3622080 2099 0 TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 2082 2101 0AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 2105 2124 0 GGCCCCGAGTGTGCTGACCA405979 5-10-5 366 2110 2129 0 GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 3672112 2131 0 CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 2114 2133 0ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 2118 2137 0 GGCCATCCGTGTAGGCCCCG405983 5-10-5 371 2120 2139 0 GTGGCCATCCGTGTAGGCCC 405984 5-10-5 3722168 2187 0 CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 2170 2189 0AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 2175 2194 0 GGAGAAACTGGAGCAGCTCA405987 5-10-5 375 2177 2196 0 CTGGAGAAACTGGAGCAGCT 405996 5-10-5 4122357 2376 0 GACTTTGCATTCCAGACCTG 405997 5-10-5 415 2362 2381 0TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 2568 2587 0 TCCGGCAGCAGATGGCAACG406025 5-10-5 216 1362 1381 0 GTGTCTAGGAGATACACCTC 406026 5-10-5 2171367 1386 0 TGCTGGTGTCTAGGAGATAC 406027 5-10-5 226 1404 1423 0ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 1406 1425 0 CCATGACCCTGCCCTCGATT406029 5-10-5 228 1408 1427 0 GACCATGACCCTGCCCTCGA 406030 5-10-5 2291412 1431 0 CGGTGACCATGACCCTGCCC 406031 5-10-5 230 1414 1433 0GTCGGTGACCATGACCCTGC 406032 5-10-5 232 1418 1437 0 CGAAGTCGGTGACCATGACC406033 5-10-5 237 1497 1516 0 CCTGCCAGGTGGGTGCCATG 406034 5-10-5 2381502 1521 0 CCACCCCTGCCAGGTGGGTG 406035 5-10-5 239 1507 1526 0GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 1519 1538 0 GGCATCCCGGCCGCTGACCA406037 5-10-5 242 1522 1541 0 GCCGGCATCCCGGCCGCTGA 406038 5-10-5 2451529 1548 0 TGGCCACGCCGGCATCCCGG 406039 5-10-5 257 1566 1585 0CAGTTGAGCACGCGCAGGCT 406040 5-10-5 258 1568 1587 0 GGCAGTTGAGCACGCGCAGG406041 5-10-5 259 1572 1591 0 CCTTGGCAGTTGAGCACGCG 406042 5-10-5 2601577 1596 0 CCTTCCCTTGGCAGTTGAGC 406043 5-10-5 261 1581 1600 0GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 262 1583 1602 0 CCGTGCCCTTCCCTTGGCAG406045 5-10-5 266 1626 1645 0 TGGCTTTTCCGAATAAACTC 408642 5-10-5 2641606 1625 0 CAGGCCTATGAGGGTGCCGC 408653 5-10-5 354 2003 2022 0TGGTCCTCAGGGAACCAGGC 409126 5-10-5 447 1534 1553 1 ACCCTTGGTCACGCCGGCAT410536 5-10-5 243 1527 1546 0 GCCACGCCGGCATCCCGGCC 410537 5-10-5 2441528 1547 0 GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 1539 1558 0CTGGCACCCTTGGCCACGCC 410539 5-10-5 255 1540 1559 0 GCTGGCACCCTTGGCCACGC410540 5-10-5 356 2005 2024 0 GCTGGTCCTCAGGGAACCAG 410562 5-10-5 4112356 2375 0 ACTTTGCATTCCAGACCTGG 410563 5-10-5 413 2358 2377 0TGACTTTGCATTCCAGACCT 410564 5-10-5 414 2359 2378 0 TTGACTTTGCATTCCAGACC410565 5-10-5 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410566 5-10-5 4202561 2580 0 GCAGATGGCAACGGCTGTCA 410567 5-10-5 421 2562 2581 0AGCAGATGGCAACGGCTGTC 410568 5-10-5 422 2563 2582 0 CAGCAGATGGCAACGGCTGT410569 5-10-5 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410570 5-10-5 4242566 2585 0 CGGCAGCAGATGGCAACGGC 410571 5-10-5 425 2567 2586 0CCGGCAGCAGATGGCAACGG 410572 5-10-5 427 2569 2588 0 CTCCGGCAGCAGATGGCAAC410573 5-10-5 428 2570 2589 0 GCTCCGGCAGCAGATGGCAA 410733 5-10-5 2311416 1435 0 AAGTCGGTGACCATGACCCT 410734 5-10-5 279 1749 1768 0CCGGCAGCGGTGACCAGCAC 410737 5-10-5 313 1839 1858 0 GCAGGTGCTGCAGTCGCTGG410738 5-10-5 324 1915 1934 0 GCTCGGCAGACAGCATCATG 410739 5-10-5 3361963 1982 0 CAGAGAAGTGGATCAGTCTC 410740 5-10-5 345 1991 2010 0AACCAGGCCTCATTGATGAC 410741 5-10-5 369 2116 2135 0 CCATCCGTGTAGGCCCCGAG410753 5-10-5 215 1351 1370 0 ATACACCTCCACCAGGCTGC 410754 5-10-5 2331428 1447 0 GGCACATTCTCGAAGTCGGT 410755 5-10-5 234 1463 1482 0TGGCCTGTCTGTGGAAGCGG 410756 5-10-5 235 1490 1509 0 GGTGGGTGCCATGACTGTCA410757 5-10-5 240 1515 1534 0 TCCCGGCCGCTGACCACCCC 410758 5-10-5 2561545 1564 0 CGCATGCTGGCACCCTTGGC 410759 5-10-5 263 1594 1613 0GGTGCCGCTAACCGTGCCCT 410760 5-10-5 265 1618 1637 0 CCGAATAAACTCCAGGCCTA410761 5-10-5 271 1649 1668 0 GTGGCCCCACAGGCTGGACC 410762 5-10-5 2721662 1681 0 AGCAGCACCACCAGTGGCCC 410763 5-10-5 273 1684 1703 0GCTGTACCCACCCGCCAGGG 410764 5-10-5 274 1730 1749 0 CGACCCCAGCCCTCGCCAGG410769 5-10-5 318 1858 1877 0 TCCCACTCTGTGACACAAAG 410770 5-10-5 3271933 1952 0 CGGCCAGGGTGAGCTCCGGC 410771 5-10-5 360 2061 2080 0ACCTGCCCCATGGGTGCTGG 410776 5-10-5 408 2245 2264 0 GTGCCAAGGTCCTCCACCTC410777 5-10-5 409 2265 2284 0 TCAGCACAGGCGGCTTGTGG 410778 5-10-5 4102295 2314 0 CCACGCACTGGTTGGGCTGA 410779 5-10-5 416 2375 2394 0CGGGATTCCATGCTCCTTGA 410780 5-10-5 417 2405 2424 0 GCAGGCCACGGTCACCTGCT410781 5-10-5 418 2442 2461 0 GGAGGGCACTGCAGCCAGTC 410782 5-10-5 4292580 2599 0 CCAGGTGCCGGCTCCGGCAG 410783 5-10-5 430 2590 2609 0GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 3. In certain such embodiments, thenucleotide sequences illustrated in Table 12 have a 3-14-3 gap-widenedmotif. Table 14 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 3, having a 3-14-3 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 14 Gapmer antisense compounds having a 3-14-3 motif targeted toSEQ ID NO: 3 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ IDIsis No Motif NO NO: 3 NO: 3 Mismatches Sequence (5′-3′) 399923 3-14-385 155 174 0 ACAAATTCCCAGACTCAGCA 399983 3-14-3 100 220 239 0ATCTCAGGACAGGTGAGCAA 399984 3-14-3 116 234 253 0 GAGTAGAGATTCTCATCTCA399924 3-14-3 129 290 309 0 GTGCCATCTGAACAGCACCT 399985 3-14-3 117 302321 0 GAGTCTTCTGAAGTGCCATC 399986 3-14-3 81 377 396 0AAGCAGGGCCTCAGGTGGAA 399925 3-14-3 110 400 419 0 CCTGGAACCCCTGCAGCCAG399926 3-14-3 152 451 470 0 TTCAGGCAGGTTGCTGCTAG 399987 3-14-3 83 460479 0 AAGGAAGACTTCAGGCAGGT 399927 3-14-3 140 472 491 0TCAGCCAGGCCAAAGGAAGA 399988 3-14-3 137 586 605 0 TAGGGAGAGCTCACAGATGC399928 3-14-3 136 596 615 0 TAGGAGAAAGTAGGGAGAGC 399929 3-14-3 132 653672 0 TAAAAGCTGCAAGAGACTCA 399930 3-14-3 139 741 760 0TCAGAGAAAACAGTCACCGA 399989 3-14-3 92 871 890 0 AGAGACAGGAAGCTGCAGCT399931 3-14-3 142 878 897 0 TCATTTTAGAGACAGGAAGC 399932 3-14-3 113 915934 0 GAATAACAGTGATGTCTGGC 399933 3-14-3 138 968 987 0TCACAGCTCACCGAGTCTGC 399934 3-14-3 98 998 1017 0 AGTGTAAAATAAAGCCCCTA399935 3-14-3 96 1075 1094 0 AGGACCCAAGTCATCCTGCT 399936 3-14-3 124 11051124 0 GGCCATCAGCTGGCAATGCT 399990 3-14-3 82 1144 1163 0AAGGAAAGGGAGGCCTAGAG 399937 3-14-3 133 1149 1168 0 TAGACAAGGAAAGGGAGGCC399938 3-14-3 103 1155 1174 0 ATTTCATAGACAAGGAAAGG 399939 3-14-3 1551275 1294 0 CTTATAGTTAACACACAGAA 399991 3-14-3 156 1283 1302 0AAGTCAACCTTATAGTTAAC 399940 3-14-3 146 1315 1334 0 TGACATTTGTGGGAGAGGAG399941 3-14-3 161 1322 1341 0 TCCAAGGTGACATTTGTGGG 399882 3-14-3 19 13651384 0 CTGGTGTCTAGGAGATACAC 399953 3-14-3 20 1370 1389 0GTATGCTGGTGTCTAGGAGA 399883 3-14-3 21 1390 1409 0 GATTTCCCGGTGGTCACTCT399954 3-14-3 22 1396 1415 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 23 14101429 0 GTGACCATGACCCTGCCCTC 399884 3-14-3 24 1420 1439 0CTCGAAGTCGGTGACCATGA 399885 3-14-3 25 1453 1472 0 GTGGAAGCGGGTCCCGTCCT399886 3-14-3 26 1500 1519 0 ACCCCTGCCAGGTGGGTGCC 399956 3-14-3 27 15051524 0 TGACCACCCCTGCCAGGTGG 399887 3-14-3 28 1534 1553 0ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 1570 1589 0 TTGGCAGTTGAGCACGCGCA399957 3-14-3 30 1575 1594 0 TTCCCTTGGCAGTTGAGCAC 399889 3-14-3 31 16071626 0 CCAGGCCTATGAGGGTGCCG 399890 3-14-3 32 1628 1647 0GCTGGCTTTTCCGAATAAAC 399891 3-14-3 33 1740 1759 0 GTGACCAGCACGACCCCAGC399897 3-14-3 39 1822 1841 0 TGGAGGCACCAATGATGTCC 399960 3-14-3 40 18331852 0 GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 1844 1863 0ACAAAGCAGGTGCTGCAGTC 399898 3-14-3 101 1898 1917 0 ATGGCTGCAATGCCAGCCAC399962 3-14-3 42 1903 1922 0 GCATCATGGCTGCAATGCCA 399963 3-14-3 43 19111930 0 GGCAGACAGCATCATGGCTG 399964 3-14-3 44 1959 1978 0GAAGTGGATCAGTCTCTGCC 399899 3-14-3 45 1967 1986 0 TTGGCAGAGAAGTGGATCAG399965 3-14-3 46 1972 1991 0 CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 19781997 0 TGATGACATCTTTGGCAGAG 399967 3-14-3 48 1985 2004 0GCCTCATTGATGACATCTTT 399968 3-14-3 49 1997 2016 0 TCAGGGAACCAGGCCTCATT399900 3-14-3 50 2002 2021 0 GGTCCTCAGGGAACCAGGCC 399901 3-14-3 87 20092028 0 ACCCGCTGGTCCTCAGGGAA 399969 3-14-3 51 2016 2035 0GGTCAGTACCCGCTGGTCCT 399902 3-14-3 119 2038 2057 0 GCAGGGCGGCCACCAGGTTG399903 3-14-3 52 2073 2092 0 AAACAGCTGCCAACCTGCCC 399970 3-14-3 53 20782097 0 CTGCAAAACAGCTGCCAACC 399942 3-14-3 104 2085 2104 0CACAGTCCTGCAAAACAGCT 399992 3-14-3 130 2095 2114 0 GTGCTGACCACACAGTCCTG399904 3-14-3 54 2108 2127 0 GTAGGCCCCGAGTGTGCTGA 399971 3-14-3 55 21732192 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 56 2179 2198 0TCCTGGAGAAACTGGAGCAG 399908 3-14-3 60 2355 2374 0 CTTTGCATTCCAGACCTGGG399973 3-14-3 61 2360 2379 0 CTTGACTTTGCATTCCAGAC 399909 3-14-3 62 25652584 0 GGCAGCAGATGGCAACGGCT 399910 3-14-3 154 2665 2684 0TTTTAAAGCTCAGCCCCAGC 399974 3-14-3 63 2670 2689 0 AACCATTTTAAAGCTCAGCC399911 3-14-3 64 2759 2778 0 TCAAGGGCCAGGCCAGCAGC 399975 3-14-3 65 27642783 0 CCCACTCAAGGGCCAGGCCA 399976 3-14-3 122 2837 2856 0GGAGGGAGCTTCCTGGCACC 399912 3-14-3 66 2852 2871 0 ATGCCCCACAGTGAGGGAGG399913 3-14-3 67 2861 2880 0 AATGGTGAAATGCCCCACAG 399914 3-14-3 153 29042923 0 TTGGGAGCAGCTGGCAGCAC 399915 3-14-3 68 3005 3024 0CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 3087 3106 0 GGAGATGAGGGCCATCAGCA399917 3-14-3 70 3155 3174 0 TAGATGCCATCCAGAAAGCT 399977 3-14-3 71 31613180 0 TCTGGCTAGATGCCATCCAG 399918 3-14-3 72 3238 3257 0GGCATAGAGCAGAGTAAAGG 399978 3-14-3 73 3243 3262 0 AGCCTGGCATAGAGCAGAGT399979 3-14-3 135 3249 3268 0 TAGCACAGCCTGGCATAGAG 399919 3-14-3 1123482 3501 0 GAAGAGGCTTGGCTTCAGAG 399980 3-14-3 74 3488 3507 0AAGTAAGAAGAGGCTTGGCT 399920 3-14-3 75 3692 3711 0 GCTCAAGGAGGGACAGTTGT399921 3-14-3 76 3727 3746 0 AAAGATAAATGTCTGCTTGC 399981 3-14-3 77 37323751 0 ACCCAAAAGATAAATGTCTG 399922 3-14-3 78 3798 3817 0TCTTCAAGTTACAAAAGCAA 399982 3-14-3 99 3805 3824 0 ATAAATATCTTCAAGTTACA405604 3-14-3 236 1493 1512 0 CCAGGTGGGTGCCATGACTG 405641 3-14-3 3732170 2189 0 AACTGGAGCAGCTCAGCAGC 410583 3-14-3 243 1527 1546 0GCCACGCCGGCATCCCGGCC 410584 3-14-3 244 1528 1547 0 GGCCACGCCGGCATCCCGGC410585 3-14-3 245 1529 1548 0 TGGCCACGCCGGCATCCCGG 410586 3-14-3 2461530 1549 0 TTGGCCACGCCGGCATCCCG 410587 3-14-3 247 1531 1550 0CTTGGCCACGCCGGCATCCC 410588 3-14-3 248 1532 1551 0 CCTTGGCCACGCCGGCATCC410589 3-14-3 249 1533 1552 0 CCCTTGGCCACGCCGGCATC 410590 3-14-3 2501535 1554 0 CACCCTTGGCCACGCCGGCA 410591 3-14-3 251 1536 1555 0GCACCCTTGGCCACGCCGGC 410592 3-14-3 252 1537 1556 0 GGCACCCTTGGCCACGCCGG410593 3-14-3 253 1538 1557 0 TGGCACCCTTGGCCACGCCG 410594 3-14-3 2541539 1558 0 CTGGCACCCTTGGCCACGCC 410595 3-14-3 255 1540 1559 0GCTGGCACCCTTGGCCACGC 410596 3-14-3 348 1995 2014 0 AGGGAACCAGGCCTCATTGA410597 3-14-3 349 1996 2015 0 CAGGGAACCAGGCCTCATTG 410598 3-14-3 3501998 2017 0 CTCAGGGAACCAGGCCTCAT 410599 3-14-3 351 1999 2018 0CCTCAGGGAACCAGGCCTCA 410600 3-14-3 352 2000 2019 0 TCCTCAGGGAACCAGGCCTC410601 3-14-3 353 2001 2020 0 GTCCTCAGGGAACCAGGCCT 410602 3-14-3 3542003 2022 0 TGGTCCTCAGGGAACCAGGC 410603 3-14-3 355 2004 2023 0CTGGTCCTCAGGGAACCAGG 410604 3-14-3 356 2005 2024 0 GCTGGTCCTCAGGGAACCAG410633 3-14-3 411 2356 2375 0 ACTTTGCATTCCAGACCTGG 410634 3-14-3 4122357 2376 0 GACTTTGCATTCCAGACCTG 410635 3-14-3 413 2358 2377 0TGACTTTGCATTCCAGACCT 410636 3-14-3 414 2359 2378 0 TTGACTTTGCATTCCAGACC410637 3-14-3 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410638 3-14-3 4202561 2580 0 GCAGATGGCAACGGCTGTCA 410639 3-14-3 421 2562 2581 0AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 2563 2582 0 CAGCAGATGGCAACGGCTGT410641 3-14-3 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410642 3-14-3 4242566 2585 0 CGGCAGCAGATGGCAACGGC 410643 3-14-3 425 2567 2586 0CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 2568 2587 0 TCCGGCAGCAGATGGCAACG410645 3-14-3 427 2569 2588 0 CTCCGGCAGCAGATGGCAAC 410646 3-14-3 4282570 2589 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gap-widened antisensecompounds are targeted to SEQ ID NO: 3. In certain such embodiments, thenucleotide sequences illustrated in Table 12 have a 2-13-5 gap-widenedmotif. Table 15 illustrates gap-widened antisense compounds targeted toSEQ ID NO: 3, having a 2-13-5 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 15 Gapmer antisense compounds having a 2-13-5 motif targeted toSEQ ID NO: 3 5′ Target 3′ Target Site to Site to SEQ ID SEQ ID SEQ IDISIS No Motiff NO NO: 3 NO: 3 Mismatches Sequences (5′-3′) 410658 2-13-5243 1527 1546 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 244 1528 1547 0GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 1529 1548 0 TGGCCACGCCGGCATCCCGG410661 2-13-5 246 1530 1549 0 TTGGCCACGCCGGCATCCCG 410662 2-13-5 2471531 1550 0 CTTGGCCACGCCGGCATCCC 410663 2-13-5 248 1532 1551 0CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 1533 1552 0 CCCTTGGCCACGCCGGCATC410665 2-13-5 28 1534 1553 0 ACCCTTGGCCACGCCGGCAT 410666 2-13-5 250 15351554 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 251 1536 1555 0GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 1537 1556 0 GGCACCCTTGGCCACGCCGG410669 2-13-5 253 1538 1557 0 TGGCACCCTTGGCCACGCCG 410670 2-13-5 2541539 1558 0 CTGGCACCCTTGGCCACGCC 410671 2-13-5 255 1540 1559 0GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 1995 2014 0 AGGGAACCAGGCCTCATTGA410673 2-13-5 349 1996 2015 0 CAGGGAACCAGGCCTCATTG 410674 2-13-5 49 19972016 0 TCAGGGAACCAGGCCTCATT 410675 2-13-5 350 1998 2017 0CTCAGGGAACCAGGCCTCAT 410676 2-13-5 351 1999 2018 0 CCTCAGGGAACCAGGCCTCA410677 2-13-5 352 2000 2019 0 TCCTCAGGGAACCAGGCCTC 410678 2-13-5 3532001 2020 0 GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 2002 2021 0GGTCCTCAGGGAACCAGGCC 410680 2-13-5 354 2003 2022 0 TGGTCCTCAGGGAACCAGGC410681 2-13-5 355 2004 2023 0 CTGGTCCTCAGGGAACCAGG 410682 2-13-5 3562005 2024 0 GCTGGTCCTCAGGGAACCAG 410713 2-13-5 60 2355 2374 0CTTTGCATTCCAGACCTGGG 410714 2-13-5 411 2356 2375 0 ACTTTGCATTCCAGACCTGG410715 2-13-5 412 2357 2376 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 4132358 2377 0 TGACTTTGCATTCCAGACCT 410717 2-13-5 414 2359 2378 0TTGACTTTGCATTCCAGACC 410718 2-13-5 61 2360 2379 0 CTTGACTTTGCATTCCAGAC410719 2-13-5 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 4202561 2580 0 GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 2562 2581 0AGCAGATGGCAACGGCTGTC 410722 2-13-5 422 2563 2582 0 CAGCAGATGGCAACGGCTGT410723 2-13-5 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 25652584 0 GGCAGCAGATGGCAACGGCT 410725 2-13-5 424 2566 2585 0CGGCAGCAGATGGCAACGGC 410726 2-13-5 425 2567 2586 0 CCGGCAGCAGATGGCAACGG410727 2-13-5 426 2568 2587 0 TCCGGCAGCAGATGGCAACG 410728 2-13-5 4272569 2588 0 CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 2570 2589 0GCTCCGGCAGCAGATGGCAA

The following embodiments set forth target regions of PCSK9 nucleicacids. Also illustrated are examples of antisense compounds targeted tothe target regions. It is understood that the sequence set forth in eachSEQ ID NO is independent of any modification to a sugar moiety, aninternucleoside linkage, or a nucleobase. As such, antisense compoundsdefined by a SEQ ID NO may comprise, independently, one or moremodifications to a sugar moiety, an internucleoside linkage, or anucleobase. Antisense compounds described by Isis Number (Isis No)indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9nucleic acid. In certain embodiment, such compounds contain at least an8 nucleotide core sequence in common. In certain embodiments, suchcompounds sharing at least an 8 nucleotide core sequence targets thefollowing nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419,451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124,1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447,1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435,1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541,1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556,1530-1557, 1531-1557, 1532-1551, 1532-1558, 1533-1559, 1534-1559,1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626,1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775,1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865,1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938,1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035,1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023,1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139,2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198,2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461,2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589,2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783,2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184,3238-3268, 3482-3711, or 3727-3751, or 3798-3824.

In certain embodiments, a target region is nucleotides 290-321 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 290-321 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 117 or 129. In certainsuch embodiments, an antisense compound targeted to nucleotides 290-321of SEQ ID NO: 3 is selected from ISIS NOs: 395202, 399924, 399829 or399985.

In certain embodiments, a target region is nucleotides 220-253 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 220-253 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 100 or 116. In certainsuch embodiments, an antisense compound targeted to nucleotides 220-253of SEQ ID NO: 3 is selected from ISIS NOs: 399827, 399983, 399828 or399984.

In certain embodiments, a target region is nucleotides 377-419 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 377-419 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain suchembodiments, an antisense compound targeted to nucleotides 377-419 ofSEQ ID NO: 3 is selected from ISIS NOs: 399830, 399986, 395203 or399925.

In certain embodiments, a target region is nucleotides 451-615 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 451-615 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 152, 140, 137 or 136. Incertain such embodiments, an antisense compound targeted to nucleotides451-615 of SEQ ID NO: 3 is selected from ISIS NOs: 395204, 399926,399831, 399987, 395205, 399927, 399832, 399988, 395206 or 399928.

In certain embodiments, a target region is nucleotides 653-672 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 653-672 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 132. In certain suchembodiments, an antisense compound targeted to nucleotides 653-672 ofSEQ ID NO: 3 is selected from ISIS NOs: 395207 or 399929.

In certain embodiments, a target region is nucleotides 741-760 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 741-760 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 139. In certain suchembodiments, an antisense compound targeted to nucleotides 741-760 ofSEQ ID NO: 3 is selected from ISIS NOs: 395208 or 399930.

In certain embodiments, a target region is nucleotides 871-897 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 871-897 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain suchembodiments, an antisense compound targeted to nucleotides 871-897 ofSEQ ID NO: 3 is selected from ISIS NOs: 399833, 399989, 395209 or399931.

In certain embodiments, a target region is nucleotides 915-934 of SEQ IDNO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 915-934 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 113. In certain suchembodiments, an antisense compound targeted to nucleotides 915-934 ofSEQ ID NO: 3 is selected from ISIS NOs: 395210 or 399932.

In certain embodiments, a target region is nucleotides 968-1017 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 968-1017 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain suchembodiments, an antisense compound targeted to nucleotides 968-1017 ofSEQ ID NO: 3 is selected from ISIS NOs: 395211, 399933, 395212, or399934.

In certain embodiments, a target region is nucleotides 1075-1124 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1075-1124 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain suchembodiments, an antisense compound targeted to nucleotides 1075-1124 ofSEQ ID NO: 3 is selected from ISIS NOs: 395213, 399935, 395214, or399936.

In certain embodiments, a target region is nucleotides 1075-1174 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1075-1174 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 82, 96, 103, 124, or 133.In certain such embodiments, an antisense compound targeted tonucleotides 1075-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 395213,399935, 395214, 399936, 399834, 399990, 395215, 399937, 395216, or399938.

In certain embodiments, a target region is nucleotides 1144-1174 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1144-1174 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 82, 133 or 103. In certainsuch embodiments, an antisense compound targeted to nucleotides1144-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 399834, 399990,395215, 399937, 395216 or 399938.

In certain embodiments, a target region is nucleotides 1275-1302 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1275-1302 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 155 or 156. In certainsuch embodiments, an antisense compound targeted to nucleotides1275-1302 of SEQ ID NO: 3 is selected from ISIS NOs: 395217, 399939,399835 or 399991.

In certain embodiments, a target region is nucleotides 1315-1341 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1315-1341 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 146 or 161. In certainsuch embodiments, an antisense compound targeted to nucleotides1315-1341 of SEQ ID NO: 3 is selected from ISIS NOs: 395218, 399940,395219 or 399941.

In certain embodiments, a target region is nucleotides 1351-1389 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1351-1389 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217 or 20.In certain such embodiments, an antisense compound targeted tonucleotides 1351-1389 of SEQ ID NO: 3 is selected from ISIS NOs: 410753,406025, 395160, 399882, 406026, 399797 or 399953.

In certain embodiments, a target region is nucleotides 1351-1447 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1351-1447 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217, 20, 21,218, 219, 220, 221, 22, 222, 223, 224, 225, 226, 227, 228, 23, 229, 230,231, 232, 24, or 233. In certain such embodiments, an antisense compoundtargeted to nucleotides 1351-1447 of SEQ ID NO: 3 is selected from ISISNOs: 410753, 406025, 395160, 399882, 406026, 399797, 399953, 395161,399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874,405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031,410733, 406032, 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 1365-1439 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1365-1439 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24,217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230,231, or 232. In certain such embodiments, an antisense compound targetedto nucleotides 1365-1439 of SEQ ID NO: 3 is selected from ISIS NOs:395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870,405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027,406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162,or 399884.

In certain embodiments, a target region is nucleotides 1390-1417 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1390-1417 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 218, 219, 220, 221,22, 222 or 223. In certain such embodiments, an antisense compoundtargeted to nucleotides 1390-1417 of SEQ ID NO: 3 is selected from ISISNOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954,405873 or 405874.

In certain embodiments, a target region is nucleotides 1390-1429 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1390-1429 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 217, 218, 219,220, 221, 222, 223, 224, 225, 226, 227, or 228. In certain suchembodiments, an antisense compound targeted to nucleotides 1390-1429 ofSEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797,399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954,405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, or399955.

In certain embodiments, a target region is nucleotides 1390-1439 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1390-1439 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 217, 218,219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, or 232.In certain such embodiments, an antisense compound targeted tonucleotides 1390-1439 of SEQ ID NO: 3 is selected from ISIS NOs: 395160,399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871,405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028,406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or399884.

In certain embodiments, a target region is nucleotides 1399-1425 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1399-1425 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. Incertain such embodiments, an antisense compound targeted to nucleotides1399-1425 of SEQ ID NO: 3 is selected from ISIS NOs: 405875, 405876,406027 or 406028.

In certain embodiments, a target region is nucleotides 1408-1435 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1408-1435 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 228, 23, 229, 230 or 231.In certain such embodiments, an antisense compound targeted tonucleotides 1408-1435 of SEQ ID NO: 3 is selected from ISIS NOs: 406029,399799, 399955, 406030, 406031 or 410733.

In certain embodiments, a target region is nucleotides 1420-1447 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1420-1447 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain suchembodiments, an antisense compound targeted to nucleotides 1420-1447 ofSEQ ID NO: 3 is selected from ISIS NOs: 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 1453-1482 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1453-1482 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain suchembodiments, an antisense compound targeted to nucleotides 1453-1482 ofSEQ ID NO: 3 is selected from ISIS NOs: 395163, 399885 or 410755.

In certain embodiments, a target region is nucleotides 1490-1516 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1490-1516 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. Incertain such embodiments, an antisense compound targeted to nucleotides1490-1516 of SEQ ID NO: 3 is selected from ISIS NOs: 410756, 405604, or406033.

In certain embodiments, a target region is nucleotides 1490-1564 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1490-1564 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 235, 236, 237, 26, 238,27, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 447, 28, 250,251, 252, 253, 254, 255, or 256. In certain such embodiments, anantisense compound targeted to nucleotides 1490-1564 of SEQ ID NO: 3 isselected from ISIS NOs: 410756, 405604, 406033, 395164, 399886, 406034,399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658,410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661,405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, or410758.

In certain embodiments, a target region is nucleotides 1500-1526 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1500-1526 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 26, 238, 27, or 239. Incertain such embodiments, an antisense compound targeted to nucleotides1500-1526 of SEQ ID NO: 3 is selected from ISIS NOs: 395164, 399886,406034, 399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 1515-1541 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1515-1541 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. Incertain such embodiments, an antisense compound targeted to nucleotides1515-1541 of SEQ ID NO: 3 is selected from ISIS NOs: 399886, 406034,399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 1527-1553 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1527-1553 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247,248, 447, 28 or 249. In certain such embodiments, an antisense compoundtargeted to nucleotides 1527-1553 of SEQ ID NO: 3 is selected from ISISNOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585,410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588,410663, 405880, 410589, 410664, 395165, 399887, or 410665.

In certain embodiments, a target region is nucleotides 1527-1554 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1527-1554 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247,248, 249, 447, 28, or 250. In certain such embodiments, an antisensecompound targeted to nucleotides 1527-1554 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038,410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879,410588, 410663, 405880, 410589, 410664, 395165, 399887, 405881, 410590,410666, or 410665.

In certain embodiments, a target region is nucleotides 1528-1554 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1528-1554 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 244, 245, 246, 247, 248,249, 447, 28, or 250. In certain such embodiments, an antisense compoundtargeted to nucleotides 1528-1554 of SEQ ID NO: 3 is selected from ISISNOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586,410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589,410664, 395165, 399887, 409126, 410665, or 405881.

In certain embodiments, a target region is nucleotides 1529-1555 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1529-1555 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249,447, 28, 250 or 251. In certain such embodiments, an antisense compoundtargeted to nucleotides 1529-1555 of SEQ ID NO: 3 is selected from ISISNOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591,410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or410667.

In certain embodiments, a target region is nucleotides 1529-1556 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1529-1556 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249,447, 28, 250, 251, or 252. In certain such embodiments, an antisensecompound targeted to nucleotides 1529-1556 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880,405881, 405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589,410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665,410666, 410667, or 410668.

In certain embodiments, a target region is nucleotides 1530-1556 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1530-1556 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 246, 247, 248, 249, 447,28, 250, 251, or 252. In certain such embodiments, an antisense compoundtargeted to nucleotides 1530-1556 of SEQ ID NO: 3 is selected from ISISNOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589, 410590,410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666,410667, 405884, 410593, 410669 or 410668.

In certain embodiments, a target region is nucleotides 1530-1557 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1530-1557 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249,250, 251, 252, 253, or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 1530-1557 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881,405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590,410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666,410667, 410668, or 410669.

In certain embodiments, a target region is nucleotides 1531-1557 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1531-1557 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250,251, 252, 253, or 447. In certain such embodiments, an antisensecompound targeted to nucleotides 1531-1557 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395165, 399887, 405878, 405879, 405880, 405881, 405882,405883, 405884, 409126, 410587, 410588, 410589, 410590, 410591, 410592,410593, 410662, 410663, 410664, 410665, 410666, 410667, 410668, or410669.

In certain embodiments, a target region is nucleotides 1532-1551 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1532-1551 of SEQ ID NO: 3. In one such embodiment, anantisense compound targeted to nucleotides 1532-1551 of SEQ ID NO: 3 isISIS NO: 405879.

In certain embodiments, a target region is nucleotides 1532-1558 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1532-1558 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251,252, 253, 254 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 1532-1558 of SEQ ID NO: 3 is selected from ISISNOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887,410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592,410668, 405884, 410593, 410669, 410538, 410594, or 410670.

In certain embodiments, a target region is nucleotides 1533-1559 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1533-1559 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252,253, 254, 255 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 1533-1559 of SEQ ID NO: 3 is selected from ISISNOs: 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590,410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593,410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1534-1559 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1534-1559 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253,254, 255 or 447. In certain such embodiments, an antisense compoundtargeted to nucleotides 1534-1559 of SEQ ID NO: 3 is selected from ISISNOs: 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591,410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594,410670, or 410671.

In certain embodiments, a target region is nucleotides 1535-1559 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1535-1559 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254,255 or 447. In certain such embodiments, an antisense compound targetedto nucleotides 1535-1559 of SEQ ID NO: 3 is selected from ISIS NOs:405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668,405884, 410593, 410669, 410538, 410594 or 410671.

In certain embodiments, a target region is nucleotides 1536-1559 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1536-1559 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or255. In certain such embodiments, an antisense compound targeted tonucleotides 1536-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405882,410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538,410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1537-1564 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1537-1564 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, 255,or 256. In certain such embodiments, an antisense compound targeted tonucleotides 1537-1564 of SEQ ID NO: 3 is selected from ISIS NOs: 405882,410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538,410594, 410670, 410539, 410595, 410671, or 410758.

In certain embodiments, a target region is nucleotides 1566-1602 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1566-1602 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258,259, 260, 261, or 262. In certain such embodiments, an antisensecompound targeted to nucleotides 1566-1602 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395166, 395167, 395168, 399801, 399888, 399889, 399890,399957, 405909, 405910, 405911, 405912, 406039, 406040, 406041, 406042,406043, or 406044.

In certain embodiments, a target region is nucleotides 1566-1681 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1566-1681 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258,259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, or 272.In certain such embodiments, an antisense compound targeted tonucleotides 1566-1681 of SEQ ID NO: 3 is selected from ISIS NOs: 395166,395167, 395168, 399801, 399888, 399889, 399890, 399957, 405909, 405910,405911, 405912, 406039, 406040, 406041, 406042, 406043, 406044, 406045,408642, 410759, 410760, 410761, or 410762.

In certain embodiments, a target region is nucleotides 1606-1626 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1606-1626 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 264 or 31. In certain suchembodiments, an antisense compound targeted to nucleotides 1606-1626 ofSEQ ID NO: 3 is selected from ISIS NOs: 408642, 395167, or 399889.

In certain embodiments, a target region is nucleotides 1618-1645 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1618-1645 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 265 or 266. In certainsuch embodiments, an antisense compound targeted to nucleotides1618-1645 of SEQ ID NO: 3 is selected from ISIS NOs: 410760 or 406045.

In certain embodiments, a target region is nucleotides 1626-1653 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1626-1653 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, or 269.In certain such embodiments, an antisense compound targeted tonucleotides 1626-1653 of SEQ ID NO: 3 is selected from ISIS NOs: 395168,399890, 405909, 405910, 405911, or 406045.

In certain embodiments, a target region is nucleotides 1684-1703 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1684-1703 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 273. In certain suchembodiments, an antisense compound targeted to nucleotides 1684-1703 ofSEQ ID NO: 3 is selected from ISIS NOs: 410763.

In certain embodiments, a target region is nucleotides 1730-1781 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1730-1781 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 33, 274, 275, 276,277, 278, 279, 280, 281, 282, 283, 284, or 285. In certain suchembodiments, an antisense compound targeted to nucleotides 1730-1781 ofSEQ ID NO: 3 is selected from ISIS NOs: 395169, 399891, 405913, 405914,405915, 405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734,or 410764.

In certain embodiments, a target region is nucleotides 1740-1767 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1740-1767 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278.In certain such embodiments, an antisense compound targeted tonucleotides 1740-1767 of SEQ ID NO: 3 is selected from ISIS NOs: 395169,399891, 405913, 405914, 405915, or 405916.

In certain embodiments, a target region is nucleotides 1749-1775 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1749-1775 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. Incertain such embodiments, an antisense compound targeted to nucleotides1749-1775 of SEQ ID NO: 3 is selected from ISIS NOs: 410734, 405917,405918 or 405919.

In certain embodiments, a target region is nucleotides 1758-1781 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1758-1781 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. Incertain such embodiments, an antisense compound targeted to nucleotides1758-1781 of SEQ ID NO: 3 is selected from ISIS NOs: 405920, 405921, or405922.

In certain embodiments, a target region is nucleotides 1820-1847 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1820-1847 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 39, 306, 307, 308, or 309.In certain such embodiments, an antisense compound targeted tonucleotides 1820-1847 of SEQ ID NO: 3 is selected from ISIS NOs: 395175,399897, 405937, 405938, 405939, or 405940.

In certain embodiments, a target region is nucleotides 1820-1877 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1820-1877 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 39, 40, 41, 306, 307, 308,309, 310, 311, 312, 313, 314, 315, 316, 317, or 318. In certain suchembodiments, an antisense compound targeted to nucleotides 1820-1877 ofSEQ ID NO: 3 is selected from ISIS NOs: 395175, 399804, 399805, 399897,399960, 399961, 405937, 405938, 405939, 405940, 405941, 405942, 405943,405944, 405945, 405946, 405947, 410737, or 410769.

In certain embodiments, a target region is nucleotides 1822-2198 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1822-2198 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309, 310,40, 311, 312, 313, 314, 315, 41, 41, 316, 317, 318, 101, 319, 42, 320,321, 322, 43, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333,334, 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343,344, 345, 346, 347, 3, or 366. In certain such embodiments, an antisensecompound targeted to nucleotides 1822-2198 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395175, 399897, 405938, 405939, 405940, 405941, 399804,399960, 405942, 405943, 410737, 405944, 405945, 399805, 399961, 405946,405947, 410769, 395176, 399898, 405948, 399806, 399962, 405949, 405950,405951, 399807, 399963, 405952, 410738, 405953, 405954, or 405979.

In certain embodiments, a target region is nucleotides 1830-1856 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1830-1856 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 40, 310, 311, or 312. Incertain such embodiments, an antisense compound targeted to nucleotides1830-1856 of SEQ ID NO: 3 is selected from ISIS NOs: 399804, 399960,405941, 405942, or 405943.

In certain embodiments, a target region is nucleotides 1839-1865 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1839-1865 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 41, 313, 314, 315, or 316.In certain such embodiments, an antisense compound targeted tonucleotides 1839-1865 of SEQ ID NO: 3 is selected from ISIS NOs: 399805,399961, 405944, 405945, 405946, or 410737.

In certain embodiments, a target region is nucleotides 1840-1867 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1840-1867 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 314, 315, 316, or 317. Incertain such embodiments, an antisense compound targeted to nucleotides1840-1867 of SEQ ID NO: 3 is selected from ISIS NOs: 399961, 405944,405945, 405946, 410737, or 405947.

In certain embodiments, a target region is nucleotides 1898-1924 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1898-1924 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. Incertain such embodiments, an antisense compound targeted to nucleotides1898-1924 of SEQ ID NO: 3 is selected from ISIS NOs: 395176, 399898,405948, 399806, 399962, 405949, or 405949.

In certain embodiments, a target region is nucleotides 1898-2035 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1898-2035 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327,328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341,342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355,356, 357, 358, or 359. In certain such embodiments, an antisensecompound targeted to nucleotides 1898-2035 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395176, 395177, 395178, 395179, 399806, 399807, 399808,399809, 399810, 399811, 399812, 399813, 399898, 399899, 399900, 399901,399962, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405885,405886, 405887, 405888, 405889, 405890, 405891, 405892, 405948, 405949,405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958,405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967,405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540,410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604,410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680,410681, 410682, 410738, 410739, 410740, or 410770.

In certain embodiments, a target region is nucleotides 1903-2127 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1903-2127 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 42, 320, 321, 322, 43,323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 44, 335,336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346,347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358,359, 51, 119, 360, or 54. In certain such embodiments, an antisensecompound targeted to nucleotides 1903-2127 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 399806, 399962, 405949, 405950, 405951, 399807, 399963,405952, 410738, 405953, 405954, 410770, 405955, 405956, 405957, 405958,405959, 405960, 405961, 399808, 399964, 405962, 410739, 405963, 395177,399899, 405964, 399809, 399965, 405965, 405966, 399810, or 399967.

In certain embodiments, a target region is nucleotides 1907-1934 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1907-1934 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 321, 322, 323, or 324.In certain such embodiments, an antisense compound targeted tonucleotides 1907-1934 of SEQ ID NO: 3 is selected from ISIS NOs: 405950,405951, 399807, 399963, 405952, or 410738.

In certain embodiments, a target region is nucleotides 1911-1938 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1911-1938 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 43, 323, 324, 325, or 326.In certain such embodiments, an antisense compound targeted tonucleotides 1911-1938 of SEQ ID NO: 3 is selected from ISIS NOs: 399807,399963, 405952, 410738, 405953, or 405954.

In certain embodiments, a target region is nucleotides 1946-1971 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1946-1971 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 328, 329, 330, or 331. Incertain such embodiments, an antisense compound targeted to nucleotides1946-1971 of SEQ ID NO: 3 is selected from ISIS NOs: 405955, 405956,405957, or 405958.

In certain embodiments, a target region is nucleotides 1954-1980 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1954-1980 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 332, 333, 334, or 335.In certain such embodiments, an antisense compound targeted tonucleotides 1954-1980 of SEQ ID NO: 3 is selected from ISIS NOs: 405959,405960, 405961, 399808, 399964, or 405962.

In certain embodiments, a target region is nucleotides 1959-2035 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1959-2035 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45,338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349,49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, or 51. Incertain such embodiments, an antisense compound targeted to nucleotides1959-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 399808, 399964,405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965,405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970,410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673,399812, 399968, 410674, or 399969.

In certain embodiments, a target region is nucleotides 1959-2057 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1959-2057 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45,338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349,49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, 51, or119. In certain such embodiments, an antisense compound targeted tonucleotides 1959-2057 of SEQ ID NO: 3 is selected from ISIS NOs: 399808,399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965,405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969,405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597,410673, 399812, 399968, 410674, or 399902.

In certain embodiments, a target region is nucleotides 1963-1988 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1963-1988 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 45, 336, 337 or 338. Incertain such embodiments, an antisense compound targeted to nucleotides1963-1988 of SEQ ID NO: 3 is selected from ISIS NOs: 410739, 405963,395177, 399899, or 405964.

In certain embodiments, a target region is nucleotides 1967-2035 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1967-2035 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 45, 338, 46, 339, 340, 47,341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353,50, 354, 355, 356, 357, 87, 358, 359 or 51. In certain such embodiments,an antisense compound targeted to nucleotides 1967-2035 of SEQ ID NO: 3is selected from ISIS NOs: 395177, 399899, 405964, 399809, 399965,405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969,405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597,410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599,or 399969.

In certain embodiments, a target region is nucleotides 1972-1999 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1972-1999 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 46, 47, 339, 340, or 341.In certain such embodiments, an antisense compound targeted tonucleotides 1972-1999 of SEQ ID NO: 3 is selected from ISIS NOs: 399809,399965, 405965, 405966, 399810, 399966, or 405967.

In certain embodiments, a target region is nucleotides 1982-2008 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1982-2008 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 48, 342, 343, or 344. Incertain such embodiments, an antisense compound targeted to nucleotides1982-2008 of SEQ ID NO: 3 is selected from ISIS NOs: 405968, 399811,399967, 405969, or 405970.

In certain embodiments, a target region is nucleotides 1991-2018 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1991-2018 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 345, 346, 347, 348,349, 350, or 351. In certain such embodiments, an antisense compoundtargeted to nucleotides 1991-2018 of SEQ ID NO: 3 is selected from ISISNOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889, 405890,410596, 410597, 410598, 410599, 410672, 410673, 410674, 410675, 410676,or 410740.

In certain embodiments, a target region is nucleotides 1993-2019 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1993-2019 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349,350, or 354. In certain such embodiments, an antisense compound targetedto nucleotides 1993-2019 of SEQ ID NO: 3 is selected from ISIS NOs:405885, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968,410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600,or 410677.

In certain embodiments, a target region is nucleotides 1995-2022 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1995-2022 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 348, 349, 350, 351, 352,353, 50, or 351 In certain such embodiments, an antisense compoundtargeted to nucleotides 1995-2022 of SEQ ID NO: 3 is selected from ISISNOs: 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968,410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600,405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, or410680.

In certain embodiments, a target region is nucleotides 1996-2023 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1996-2023 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351,352, 353, 354, or 355. In certain such embodiments, an antisensecompound targeted to nucleotides 1996-2023 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395178, 399812, 399900, 399968, 405888, 405889, 405890,405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600, 410601,410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678, 410679,410680, or 410681.

In certain embodiments, a target region is nucleotides 1997-2024 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1997-2024 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352,353, 354, 355, or 356. In certain such embodiments, an antisensecompound targeted to nucleotides 1997-2024 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890,410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178,399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540,410604, or 410682.

In certain embodiments, a target region is nucleotides 1998-2025 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1998-2025 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353,354, 355, 356 or 357. In certain such embodiments, an antisense compoundtargeted to nucleotides 1998-2025 of SEQ ID NO: 3 is selected from ISISNOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600,410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602,410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1999-2025 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 1999-2025 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354,355, 356, or 357. In certain such embodiments, an antisense compoundtargeted to nucleotides 1999-2025 of SEQ ID NO: 3 is selected from ISISNOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600,410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602,410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 2000-2025 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2000-2025 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 351, 352, 353, 354, 355,356, or 357. In certain such embodiments, an antisense compound targetedto nucleotides 2000-2025 of SEQ ID NO: 3 is selected from ISIS NOs:405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678,395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681,410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 2009-2035 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2009-2035 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 51, 87, 358, or 359. Incertain such embodiments, an antisense compound targeted to nucleotides2009-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395179, 399813,399901, 399969, 405973, or 405974.

In certain embodiments, a target region is nucleotides 2038-2139 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2038-2139 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 119, 130,360, 361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certainsuch embodiments, an antisense compound targeted to nucleotides2038-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 395180, 395181,395182, 395220, 399814, 399836, 399902, 399903, 399904, 399942, 399970,399992, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982,405983, 410741, or 410771.

In certain embodiments, a target region is nucleotides 2061-2139 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2061-2139 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 130, 360,361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certain suchembodiments, an antisense compound targeted to nucleotides 2061-2139 ofSEQ ID NO: 3 is selected from ISIS NOs: 395181, 395182, 395220, 399814,399836, 399903, 399904, 399942, 399970, 399992, 405975, 405976, 405977,405978, 405979, 405980, 405981, 405982, 405983, 410741, or 410771.

In certain embodiments, a target region is nucleotides 2073-2099 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2073-2099 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. Incertain such embodiments, an antisense compound targeted to nucleotides2073-2099 of SEQ ID NO: 3 is selected from ISIS NOs: 395181, 399814,399903, 399970, 405975, or 405976.

In certain embodiments, a target region is nucleotides 2078-2104 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2078-2104 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 53, 104, 362, or 363. Incertain such embodiments, an antisense compound targeted to nucleotides2078-2104 of SEQ ID NO: 3 is selected from ISIS NOs: 395220, 399814,399942, 399970, 405976, or 405977.

In certain embodiments, a target region is nucleotides 2105-2131 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2105-2131 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. Incertain such embodiments, an antisense compound targeted to nucleotides2105-2131 of SEQ ID NO: 3 is selected from ISIS NOs: 395182, 399904,405978, 405979, or 405980.

In certain embodiments, a target region is nucleotides 2112-2139 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2112-2139 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 367, 368, 369, 370, or371. In certain such embodiments, an antisense compound targeted tonucleotides 2112-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 405980,405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 2168-2198 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2168-2198 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or375. In certain such embodiments, an antisense compound targeted tonucleotides 2168-2198 of SEQ ID NO: 3 is selected from ISIS NOs: 399815,399816, 399971, 399972, 405641, 405984, 405985, 405986, or 405987.

In certain embodiments, a target region is nucleotides 2170-2177 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2170-2177 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 55, 373, 374, or 375. Incertain such embodiments, an antisense compound targeted to nucleotides2170-2177 of SEQ ID NO: 3 is selected from ISIS NOs: 399815, 399971,405641, 405985, 405986, or 405987.

In certain embodiments, a target region is nucleotides 2245-2284 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2245-2284 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 408 or 409. In certainsuch embodiments, an antisense compound targeted to nucleotides2245-2284 of SEQ ID NO: 3 is selected from ISIS NOs: 410776 or 410777.

In certain embodiments, a target region is nucleotides 2295-2394 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2295-2394 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412,413, 414, 415, or 416. In certain such embodiments, an antisensecompound targeted to nucleotides 2295-2394 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562,410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715,410716, 410717, 410718, 410778, or 410779.

In certain embodiments, a target region is nucleotides 2355-2381 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2355-2381 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413,414, or 415. In certain such embodiments, an antisense compound targetedto nucleotides 2355-2381 of SEQ ID NO: 3 is selected from ISIS NOs:395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564,410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717,or 410718.

In certain embodiments, a target region is nucleotides 2355-2394 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2355-2394 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413,414, 415, or 416. In certain such embodiments, an antisense compoundtargeted to nucleotides 2355-2394 of SEQ ID NO: 3 is selected from ISISNOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563,410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716,410717, 410718, or 410779.

In certain embodiments, a target region is nucleotides 2405-2461 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2405-2461 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 417 or 418. In certainsuch embodiments, an antisense compound targeted to nucleotides2405-2461 of SEQ ID NO: 3 is selected from ISIS NOs: 410780 or 410781.

In certain embodiments, a target region is nucleotides 2560-2587 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2560-2587 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425, or 426. In certain such embodiments, an antisensecompound targeted to nucleotides 2560-2587 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568,410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642,410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725,410726, or 410727.

In certain embodiments, a target region is nucleotides 2560-2609 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2560-2609 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422,423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments,an antisense compound targeted to nucleotides 2560-2609 of SEQ ID NO: 3is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566,410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638,410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719,410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728,or 410783.

In certain embodiments, a target region is nucleotides 2561-2588 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2561-2588 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 420, 421, 422, 423, 424,425, 426, 427, or 62. In certain such embodiments, an antisense compoundtargeted to nucleotides 2561-2588 of SEQ ID NO: 3 is selected from ISISNOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640,410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642,410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572,410645, or 410728.

In certain embodiments, a target region is nucleotides 2562-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2562-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424,425, 426, 427, or 428. In certain such embodiments, an antisensecompound targeted to nucleotides 2562-2589 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570,410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644,410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727,410728, or 410729.

In certain embodiments, a target region is nucleotides 2563-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2563-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 422, 423, 424, 425, 426,427, 428 or 62. In certain such embodiments, an antisense compoundtargeted to nucleotides 2563-2589 of SEQ ID NO: 3 is selected from ISISNOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909,410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998,410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2564-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2564-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 423, 424, 425, 426, 427,428 or 62. In certain such embodiments, an antisense compound targetedto nucleotides 2564-2589 of SEQ ID NO: 3 is selected from ISIS NOs:410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725,410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645,410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2565-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2565-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, 428 or62. In certain such embodiments, an antisense compound targeted tonucleotides 2565-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 395187,399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988,405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or410729.

In certain embodiments, a target region is nucleotides 2566-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2566-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, or428. In certain such embodiments, an antisense compound targeted tonucleotides 2566-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410570,410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727,410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2567-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2567-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 425, 426, 427, or 428. Incertain such embodiments, an antisense compound targeted to nucleotides2567-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410571, 410643,410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573,410646, or 410729.

In certain embodiments, a target region is nucleotides 2568-2589 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2568-2589 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. Incertain such embodiments, an antisense compound targeted to nucleotides2568-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 405988, 405998,410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2665-2689 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2665-2689 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain suchembodiments, an antisense compound targeted to nucleotides 2665-2689 ofSEQ ID NO: 3 is selected from ISIS NOs: 395188, 399910, 399818, or399974.

In certain embodiments, a target region is nucleotides 2759-2783 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2759-2783 SEQ ID NO: 3. In certain embodiments, an antisensecompound targeted to a PCSK9 nucleic acid comprises a nucleotidesequence selected from SEQ ID NOs: 64 or 65. In certain suchembodiments, an antisense compound targeted to nucleotides 2759-2783 ofSEQ ID NO: 3 is selected from ISIS NOs: 395189, 399911, 399819, or399975.

In certain embodiments, a target region is nucleotides 2837-2880 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2837-2880 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 66, 67, or 122. In certainsuch embodiments, an antisense compound targeted to nucleotides2837-2880 of SEQ ID NO: 3 is selected from ISIS NOs: 399820, 399976,395190, 399912, 395191, or 399913.

In certain embodiments, a target region is nucleotides 2904-2923 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 2904-2923 SEQ ID NO: 3. In certain embodiments, an antisensecompound targeted to a PCSK9 nucleic acid comprises a nucleotidesequence selected from SEQ ID NOs: 153. In certain such embodiments, anantisense compound targeted to nucleotides 2904-2923 of SEQ ID NO: 3 isselected from ISIS NOs: 395192 or 399914.

In certain embodiments, a target region is nucleotides 3005-3024 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3005-3024 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 68. In certain suchembodiments, an antisense compound targeted to nucleotides 3005-3024 ofSEQ ID NO: 3 is selected from ISIS NOs: 395193 or 399915.

In certain embodiments, a target region is nucleotides 3005-3174 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3005-3174 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 68, 431, 432, 433, 434,69, 435, 436, 437, 438 or 70. In certain such embodiments, an antisensecompound targeted to nucleotides 3005-3174 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395193, 399915, 405893, 405894, 405895, 405896, 395194,399916, 405897, 405898, 405899, 405900, 395195, or 399917.

In certain embodiments, a target region is nucleotides 3083-3110 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3083-3110 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 69, 69, 431, 432, 433,434, 435, 436, 437, or 438. In certain such embodiments, an antisensecompound targeted to nucleotides 3083-3110 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897,405898, 405899, or 405900.

In certain embodiments, a target region is nucleotides 3155-3184 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3155-3184 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441,442, 443, 444, 445, or 446. In certain such embodiments, an antisensecompound targeted to nucleotides 3155-3184 of SEQ ID NO: 3 is selectedfrom ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903,405904, 405905, 405906, 405907, or 405908.

In certain embodiments, a target region is nucleotides 3238-3268 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3238-3268 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 72, 73, or 135. In certainsuch embodiments, an antisense compound targeted to nucleotides3238-3268 of SEQ ID NO: 3 is selected from ISIS NOs: 395196, 399822,399823, 399918, 399978, or 399979.

In certain embodiments, a target region is nucleotides 3482-3711 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3482-3711 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 74, 75, or 112. In certainsuch embodiments, an antisense compound targeted to nucleotides3482-3711 of SEQ ID NO: 3 is selected from ISIS NOs: 395197, 395198,399824, 399919, 399920, or 399980.

In certain embodiments, a target region is nucleotides 3727-3751 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3727-3751 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain suchembodiments, an antisense compound targeted to nucleotides 3727-3751 ofSEQ ID NO: 3 is selected from ISIS NOs: 395199, 399921, 399825, or399981.

In certain embodiments, a target region is nucleotides 3798-3824 of SEQID NO: 3. In certain embodiments, an antisense compound is targeted tonucleotides 3798-3824 of SEQ ID NO: 3. In certain embodiments, anantisense compound targeted to a PCSK9 nucleic acid comprises anucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain suchembodiments, an antisense compound targeted to nucleotides 3798-3824 ofSEQ ID NO: 3 is selected from ISIS NOs: 395200, 399922, 399826, or399982.

In certain embodiments, short gapmer antisense compounds are targeted toa PCSK9 nucleic acid. In certain such embodiments, gapmer antisensecompounds are targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO:3. In certain such embodiments, the nucleotide sequences illustrated inTable 15.1 have a 2-10-2 gapmer motif. Table 15.1 illustrates gapmerantisense compounds targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ IDNO: 3, having a 2-10-2 motif, where the gap segment comprises2′-deoxynucleotides and each wing segment comprises nucleotidescomprising a 2′-O-methoxyethyl sugar modification. Internucleosidelinkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 15.1 Gapmer antisense compounds having a 2-10-2 motif targeted toSEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3 Target 5′ Target Site3′ Target Site SEQ ID on SEQ ID on SEQ ID SEQ NO ISIS NO Motif OligoSeqNO: 1 NO: 1 ID NO 1 406539 2-10-2 GCCTATGAGGGTGC 1079 1092 458 1 4065402-10-2 TCCAGGCCTATGAG 1084 1097 459 1 406551 2-10-2 CAGCTCAGCAGCTC 17351748 460 1 406595 2-10-2 TTAATCAGGGAGCC 2877 2890 461 2 406551 2-10-2CAGCTCAGCAGCTC 21181 21194 460 2 406595 2-10-2 TTAATCAGGGAGCC 2668426697 461 3 406539 2-10-2 GCCTATGAGGGTGC 1609 1622 458 3 406540 2-10-2TCCAGGCCTATGAG 1614 1627 459 3 406551 2-10-2 CAGCTCAGCAGCTC 2168 2181460 3 406595 2-10-2 TTAATCAGGGAGCC 3132 3145 461

In a certain embodiment, a short antisense compound has increasedpotency compared to a 20 nucleotide compound. In a certain embodimentsuch short antisense compound and 20 nucleotide compound target the sameregion. In a certain embodiment, a short antisense compound hasincreased potency compared to a 20 nucleotide compound containing thesequence of the short antisense compound.

In certain embodiments, the desired effect is a reduction in mRNA targetnucleic acid levels. In other embodiments, the desired effect isreduction of levels of protein encoded by the target nucleic acid or aphenotypic change associated with the target nucleic acid.

In one embodiment, a target region is a structurally defined region ofthe nucleic acid. For example, a target region may encompass a 3′ UTR, a5′ UTR, an exon, an intron, a coding region, a translation initiationregion, translation termination region, or other defined nucleic acidregion. In other embodiments, a target region may encompass the sequencefrom a 5′ target site of one target segment within the target region toa 3′ target site of another target segment within the target region.

Targeting includes determination of at least one target segment to whichan antisense compound hybridizes, such that a desired effect occurs. Atarget region may contain one or more target segments. Multiple targetsegments within a target region may be overlapping. Alternatively, theymay be non-overlapping. In one embodiment, target segments within atarget region are separated by no more than about 10 nucleotides on thetarget nucleic acid. In another embodiment, target segments within atarget region are separated by no more than about 5 nucleotides on thetarget nucleic acid. In additional embodiments, target segments arecontiguous.

Suitable target segments may be found within a 5′ UTR, a coding region,a 3′ UTR, an intron, or an exon. Target segments containing a startcodon or a stop codon are also suitable target segments.

The determination of suitable target segments may include a comparisonof the sequence of a target nucleic acid to other sequences throughoutthe genome. For example, the BLAST algorithm may be used to identifyregions of similarity amongst different nucleic acids. This comparisoncan prevent the selection of antisense compound sequences that mayhybridize in a non-specific manner to sequences other than a selectedtarget nucleic acid (i.e., non-target or off-target sequences).

There may be variation in activity (e.g., as defined by percentreduction of target nucleic acid levels) of the antisense compoundswithin an active target region. In one embodiment, reductions in PCSK9mRNA levels are indicative of inhibition of PCSK9 expression. Reductionsin levels of a PCSK9 protein are also indicative of inhibition of targetmRNA expression. Further, phenotypic changes are indicative ofinhibition of PCSK9 expression. For example, a decrease in LDL-C levelsis indicative of inhibition of PCSK9 expression.

Hybridization

For example, hybridization may occur between an antisense compounddisclosed herein and a PCSK9 nucleic acid. The most common mechanism ofhybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteenor reversed Hoogsteen hydrogen bonding) between complementarynucleobases of the nucleic acid molecules.

Hybridization can occur under varying conditions. Stringent conditionsare sequence-dependent and are determined by the nature and compositionof the nucleic acid molecules to be hybridized.

Methods of determining whether a sequence is specifically hybridizableto a target nucleic acid are well known in the art. In one embodiment,the antisense compounds provided herein are specifically hybridizablewith a PCSK9 nucleic acid.

The PCSK9 sequences targeted by some of the preferred antisensecompounds disclosed in Table 1 are shown in FIG. 1. The antisensecompounds are labeled with the corresponding Isis number. The bar inFIG. 1 labeled “NM_174936.2” displays the relative position of thetarget sequences within GENBANK® Accession No. NM_174936.2, firstdeposited with GENBANK® on Jun. 1, 2003 (SEQ ID NO: 1). The bar belowthat shows the corresponding structural elements of the PCSK9 mRNA,where “CDS” is the protein-encoding region of the mRNA and “UTRs” arethe 5′ and 3′ untranslated regions. As shown in FIG. 1, the sequencestargeted by the present antisense compounds are found in theprotein-encoding region of the PCSK9 mRNA.

Complementarity

An antisense compound and a target nucleic acid are complementary toeach other when a sufficient number of nucleobases of the antisensecompound can hydrogen bond with the corresponding nucleobases of thetarget nucleic acid, such that a desired effect will occur (e.g.,antisense inhibition of a target nucleic acid, such as a PCSK9 nucleicacid).

Non-complementary nucleobases between an antisense compound and a PCSK9nucleic acid may be tolerated provided that the antisense compoundremains able to specifically hybridize to a target nucleic acid.Moreover, an antisense compound may hybridize over one or more segmentsof a PCSK9 nucleic acid such that intervening or adjacent segments arenot involved in the hybridization event (e.g., a loop structure,mismatch or hairpin structure).

In some embodiments, the antisense compounds provided herein are atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98% or at least 99%complementary to a PCSK9 nucleic acid. Percent complementarity of anantisense compound with a target nucleic acid can be determined usingroutine methods.

In other embodiments, the antisense compounds provided herein are fullycomplementary (i.e, 100% complementary) to a target nucleic acid. Forexample, antisense compound may be fully complementary to a PCSK9nucleic acid. As used herein, “fully complementary” means eachnucleobase of an antisense compound is capable of precise base pairingwith the corresponding nucleobases of a target nucleic acid.

The location of a non-complementary nucleobase may be at the 5′ end or3′ end of the antisense compound. Alternatively, the non-complementarynucleobase or nucleobases may be at an internal position of theantisense compound. When two or more non-complementary nucleobases arepresent, they may be contiguous (i.e., linked) or non-contiguous. In oneembodiment, a non-complementary nucleobase is located in the wingsegment of a gapmer antisense oligonucleotide.

In one embodiment, antisense compounds up to 20 nucleobases in lengthcomprise no more than 4, no more than 3, no more than 2 or no more than1 non-complementary nucleobase(s) relative to a target nucleic acid,such as a PCSK9 nucleic acid.

In another embodiment, antisense compounds up to 30 nucleobases inlength comprise no more than 6, no more than 5, no more than 4, no morethan 3, no more than 2 or no more than 1 non-complementary nucleobase(s)relative to a target nucleic acid, such as a PCSK9 nucleic acid.

The antisense compounds provided herein also include those which arecomplementary to a portion of a target nucleic acid. As used herein,“portion” refers to a defined number of contiguous (i.e., linked)nucleobases within a region or segment of a target nucleic acid. A“portion” can also refer to a defined number of contiguous nucleobasesof an antisense compound. In one embodiment, the antisense compounds arecomplementary to at least an 8 nucleobase portion of a target segment.In another embodiment, the antisense compounds are complementary to atleast a 12 nucleobase portion of a target segment. In yet anotherembodiment, the antisense compounds are complementary to at least a 15nucleobase portion of a target segment.

Identity

The antisense compounds provided herein may also have a defined percentidentity to a particular nucleotide sequence, SEQ ID NO, or compoundrepresented by a specific Isis number. As used herein, an antisensecompound is identical to the sequence disclosed herein if it has thesame nucleobase pairing ability. For example, a RNA which containsuracil in place of thymidine in a disclosed DNA sequence would beconsidered identical to the DNA sequence since both uracil and thymidinepair with adenine. Shortened and lengthened versions of the antisensecompounds described herein as well as compounds having non-identicalbases relative to the antisense compounds provided herein also arecontemplated. The non-identical bases may be adjacent to each other ordispersed throughout the antisense compound. Percent identity of anantisense compound is calculated according to the number of bases thathave identical base pairing relative to the sequence to which it isbeing compared.

In one embodiment, the antisense compounds are at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95% or 100%identical to one or more of the antisense compounds disclosed herein.

Modifications

A nucleoside is a base-sugar combination. The nucleobase (also known asbase) portion of the nucleoside is normally a heterocyclic base moiety.Nucleotides are nucleosides that further include a phosphate groupcovalently linked to the sugar portion of the nucleoside. For thosenucleosides that include a pentofuranosyl sugar, the phosphate group canbe linked to the 2′, 3′ or 5′ hydroxyl moiety of the sugar.Oligonucleotides are formed through the covalent linkage of adjacentnucleosides to one another, to form a linear polymeric oligonucleotide.Within the oligonucleotide structure, the phosphate groups are commonlyreferred to as forming the internucleoside linkages of theoligonucleotide.

Modifications to antisense compounds encompass substitutions or changesto internucleoside linkages, sugar moieties, or nucleobases. Modifiedantisense compounds are often preferred over native forms because ofdesirable properties such as, for example, enhanced cellular uptake,enhanced affinity for nucleic acid target, increased stability in thepresence of nucleases, or increased inhibitory activity.

Chemically modified nucleosides may also be employed to increase thebinding affinity of a shortened or truncated antisense oligonucleotidefor its target nucleic acid. Consequently, comparable results can oftenbe obtained with shorter antisense compounds that have such chemicallymodified nucleosides.

Modified Internucleoside Linkages

The naturally occurring internucleoside linkage of RNA and DNA is a 3′to 5′ phosphodiester linkage. Antisense compounds having one or moremodified, i.e., non-naturally occurring, internucleoside linkages areoften selected over antisense compounds having naturally occurringinternucleoside linkages because of desirable properties such as, forexample, enhanced cellular uptake, enhanced affinity for target nucleicacids, and increased stability in the presence of nucleases.

Oligonucleotides having modified internucleoside linkages includeinternucleoside linkages that retain a phosphorus atom as well asinternucleoside linkages that do not have a phosphorus atom.Representative phosphorus containing internucleoside linkages include,but are not limited to, phosphodiesters, phosphotriesters,methylphosphonates, phosphoramidate, and phosphorothioates. Methods ofpreparation of phosphorous-containing and non-phosphorous-containinglinkages are well known.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acidcomprise one or more modified internucleoside linkages. In someembodiments, the modified internucleoside linkages are phosphorothioatelinkages. In other embodiments, each internucleoside linkage of anantisense compound is a phosphorothioate internucleoside linkage.

Modified Sugar Moieties

Antisense compounds targeted to a PCSK9 nucleic acid may contain one ormore nucleotides having modified sugar moieties. Sugar modifications mayimpart nuclease stability, binding affinity or some other beneficialbiological property to the antisense compounds. The furanosyl sugar ringof a nucleoside can be modified in a number of ways including, but notlimited to: addition of a substituent group, particularly at the 2′position; bridging of two non-geminal ring atoms to form a bicyclicnucleic acid (BNA); and substitution of an atom or group such as —S—,—N(R)— or —C(R₁)(R₂) (R═H, C1-C12 alkyl or a protecting group) for thering oxygen at the 4′-position and combinations of these such as forexample a 2′-F-5′-methyl substituted nucleoside (see WO 2008/101157,published Aug. 21, 2008, for other disclosed 5′,2′-bis substitutednucleosides) or replacement of the ribosyl ring oxygen atom with S withfurther substitution at the 2′-position (see U.S. Published ApplicationNo. 2005/0130923, published on Jun. 16, 2005) or alternatively5′-substitution of a BNA (see WO 2007/134181, published Nov. 22, 2007,wherein LNA is substituted with for example a 5′-methyl or a 5′-vinylgroup).

Modified sugars include, but are not limited to: substituted sugars,especially 2′-substituted sugars having a 2′-F, 2′-OCH₃ (2′-OMe) or a2′-O(CH₂)₂—OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituent group; andbicyclic modified sugars (BNAs), having a 4′-(CH₂)_(n)—O-2′ bridge,where n=1 or n=2, including α-L-methyleneoxy (4′-CH₂—O-2′ or4′-α-L-(CH2)-O-2′ (α-L-LNA)) BNA, β-D-methyleneoxy (4′-CH2-O-2′ or4′-β-D-(CH2)-O-2′ (β-D-LNA)) BNA and ethyleneoxy (4′-(CH2)₂—O-2′ (ENA))BNA. BNAs also include 4′-(CH2)-S-2′ and 4′-C(CH3)2-O-2′ (seeInternational Application No. PCT/US2008/068922); 4′-CH(CH3)-O-2′ and4′-CH(CH2OCH3)-O-2′ (see U.S. Pat. No. 7,399,845, issued Jul. 15, 2008);4′-CH2-N(OCH3)-2′ (see International Application No. PCT/US2008/064591);4′-CH2-O—N(CH3)-2′ (see U.S. Published Application No. 2004/0171570,published Sep. 2, 2004); 4′-CH2-N(R)—O-2′ (see U.S. Pat. No. 7,427,672,issued Sep. 23, 2008); 4′-CH2-C(CH3)-2′ and 4′-CH2-C(═CH2)-2′ (seeInternational Application No. PCT/US2008/066154); and wherein R is,independently, H, C1-C12 alkyl, or a protecting group. Bicyclic modifiedsugars also include (6'S)-6′ methyl BNA, aminooxy (4′-CH2-O—N(R)-2′)BNA, oxyamino (4′-CH2-N(R)—O-2′) BNA wherein R is, independently, H, aprotecting group or C1-C12 alkyl. Examples of nucleosides havingmodified sugar moieties also include without limitation nucleosidescomprising 5′-vinyl, 5′-methyl (R or S) and 4′-S.

The substituent at the 2′ position can also be selected from allyl,amino, azido, thio, O-allyl, O—C1-C10 alkyl, OCF₃, O(CH₂)₂SCH₃,O(CH₂)₂—O—N(Rm)(Rn), and O—CH₂—C(═O)—N(Rm)(Rn), where each Rm and Rn is,independently, H or substituted or unsubstituted C1-C10 alkyl.

In certain embodiments, such BNA modified nucleotides are high-affinitynucleotides and their incorporation into antisense compounds allows forincreased potency and improved therapeutic index. Methods for thepreparations of modified sugars are well known to those skilled in theart.

In certain embodiments, nucleosides are modified by substitution of theribosyl ring with a sugar surrogate. Such modification includes withoutlimitation, replacement of the ribosyl ring with a surrogate ring system(sometimes referred to as DNA analogs) such as a morpholino ring, acyclohexenyl ring, a cyclohexyl ring or a tetrahydropyranyl ring such asone having one of the formula:

Many other bicyclo and tricyclo sugar surrogate ring systems are alsoknow in the art that can be used to modify nucleosides for incorporationinto antisense compounds. Such ring systems can undergo variousadditional substitutions to enhance activity.

Methods for the preparations of modified sugars are well known to thoseskilled in the art.

In nucleotides having modified sugar moieties (including substitution ofa sugar surrogate), the nucleobase moieties (natural, modified or acombination thereof) are maintained for hybridization with anappropriate nucleic acid target.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acidcomprise one or more nucleotides having modified sugar moieties. In asuitable embodiment, the modified sugar moiety is 2′-MOE. In otherembodiments, the 2′-MOE modified nucleotides are arranged in a gapmermotif.

Modified Nucleobases

Nucleobase (or base) modifications or substitutions are structurallydistinguishable from, yet functionally interchangeable with, naturallyoccurring or synthetic unmodified nucleobases. Both natural and modifiednucleobases are capable of participating in hydrogen bonding. Suchnucleobase modifications may impart nuclease stability, binding affinityor some other beneficial biological property to antisense compounds.Modified nucleobases include synthetic and natural nucleobases such as,for example, 5-methylcytosine (5-me-C). Certain nucleobasesubstitutions, including 5-methylcytosine substitutions, areparticularly useful for increasing the binding affinity of an antisensecompound for a target nucleic acid. For example, 5-methylcytosinesubstitutions have been shown to increase nucleic acid duplex stabilityby 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., eds.,Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp.276-278).

Additional unmodified nucleobases include 5-hydroxymethyl cytosine,xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkylderivatives of adenine and guanine, 2-propyl and other alkyl derivativesof adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine,5-halouracil and cytosine, 5-propynyl (—C≡C—CH₃) uracil and cytosine andother alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosineand thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino,8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines andguanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other5-substituted uracils and cytosines, 7-methylguanine and7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and3-deazaadenine.

Heterocyclic base moieties may also include those in which the purine orpyrimidine base is replaced with other heterocycles, for example7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone.Nucleobases that are particularly useful for increasing the bindingaffinity of antisense compounds include 5-substituted pyrimidines,6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acidcomprise one or more modified nucleobases. In an additional embodiment,gap-widened antisense oligonucleotides targeted to a PCSK9 nucleic acidcomprise one or more modified nucleobases. In some embodiments, themodified nucleobase is 5-methylcytosine. In further embodiments, eachcytosine is a 5-methylcytosine.

Compositions and Methods for Formulating Pharmaceutical Compositions

Antisense oligonucleotides may be admixed with pharmaceuticallyacceptable active and/or inert substances for the preparation ofpharmaceutical compositions or formulations. Compositions and methodsfor the formulation of pharmaceutical compositions are dependent upon anumber of criteria, including, but not limited to, route ofadministration, extent of disease, or dose to be administered.

Antisense compound targeted to a PCSK9 nucleic acid can be utilized inpharmaceutical compositions by combining the antisense compound with asuitable pharmaceutically acceptable diluent or carrier. Apharmaceutically acceptable diluent includes phosphate-buffered saline(PBS). PBS is a diluent suitable for use in compositions to be deliveredparenterally. Accordingly, in one embodiment, employed in the methodsdescribed herein is a pharmaceutical composition comprising an antisensecompound targeted to a PCSK9 nucleic acid and a pharmaceuticallyacceptable diluent. In one embodiment, the pharmaceutically acceptablediluent is PBS. In other embodiments, the antisense compound is anantisense oligonucleotide.

Pharmaceutical compositions comprising antisense compounds encompass anypharmaceutically acceptable salts, esters, or salts of such esters, orany other oligonucleotide which, upon administration to an animal,including a human, is capable of providing (directly or indirectly) thebiologically active metabolite or residue thereof. Accordingly, forexample, the disclosure is also drawn to pharmaceutically acceptablesalts of antisense compounds, prodrugs, pharmaceutically acceptablesalts of such prodrugs, and other bioequivalents. Suitablepharmaceutically acceptable salts include, but are not limited to,sodium and potassium salts.

A prodrug can include the incorporation of additional nucleosides at oneor both ends of an antisense compound which are cleaved by endogenousnucleases within the body, to form the active antisense compound.

In certain embodiments, pharmaceutical compositions of the presentinvention comprise one or more oligonucleotides and one or moreexcipients. In certain such embodiments, excipients are selected fromwater, salt solutions, alcohol, polyethylene glycols, gelatin, lactose,amylase, magnesium stearate, talc, silicic acid, viscous paraffin,hydroxymethylcellulosem and polyvinylpyrrolidone.

In certain embodiments, a pharmaceutical composition of the presentinvention is prepared using known techniques, including, but not limitedto mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or tabletting processes.

In certain embodiments, a pharmaceutical composition of the presentinvention is a liquid (e.g., a suspension, elixir and/or solution). Incertain of such embodiments, a liquid pharmaceutical composition isprepared using ingredients known in the art, including, but not limitedto, water, glycols, oils, alcohols, flavoring agents, preservatives, andcoloring agents.

In certain embodiments, a pharmaceutical composition of the presentinvention is a solid (e.g., a powder, tablet, and/or capsule). Incertain of such embodiments, a solid pharmaceutical compositioncomprising one or more oligonucleotides is prepared using ingredientsknown in the art, including, but not limited to, starches, sugars,diluents, granulating agents, lubricants, binders, and disintegratingagents.

In certain embodiments, a pharmaceutical composition of the presentinvention is formulated as a depot preparation. Certain such depotpreparations are typically longer acting than non-depot preparations. Incertain embodiments, such preparations are administered by implantation(for example subcutaneously or intramuscularly) or by intramuscularinjection. In certain embodiments, depot preparations are prepared usingsuitable polymeric or hydrophobic materials (for example an emulsion inan acceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a delivery system. Examples of delivery systemsinclude, but are not limited to, liposomes and emulsions. Certaindelivery systems are useful for preparing certain pharmaceuticalcompositions including those comprising hydrophobic compounds. Incertain embodiments, certain organic solvents such as dimethylsulfoxideare used.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises one or more tissue-specific delivery moleculesdesigned to deliver the one or more pharmaceutical agents of the presentinvention to specific tissues or cell types. For example, in certainembodiments, pharmaceutical compositions include liposomes coated with atissue-specific antibody.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a co-solvent system. Certain of such co-solventsystems comprise, for example, benzyl alcohol, a nonpolar surfactant, awater-miscible organic polymer, and an aqueous phase. In certainembodiments, such co-solvent systems are used for hydrophobic compounds.A non-limiting example of such a co-solvent system is the VPD co-solventsystem, which is a solution of absolute ethanol comprising 3% w/v benzylalcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™, and 65% w/vpolyethylene glycol 300. The proportions of such co-solvent systems maybe varied considerably without significantly altering their solubilityand toxicity characteristics. Furthermore, the identity of co-solventcomponents may be varied: for example, other surfactants may be usedinstead of Polysorbate 80™; the fraction size of polyethylene glycol maybe varied; other biocompatible polymers may replace polyethylene glycol,e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides maysubstitute for dextrose.

In certain embodiments, a pharmaceutical composition of the presentinvention comprises a sustained-release system. A non-limiting exampleof such a sustained-release system is a semi-permeable matrix of solidhydrophobic polymers. In certain embodiments, sustained-release systemsmay, depending on their chemical nature, release pharmaceutical agentsover a period of hours, days, weeks or months.

In certain embodiments, a pharmaceutical composition of the presentinvention is prepared for oral administration. In certain of suchembodiments, a pharmaceutical composition is formulated by combining oneor more oligonucleotides with one or more pharmaceutically acceptablecarriers. Certain of such carriers enable pharmaceutical compositions tobe formulated as tablets, pills, dragees, capsules, liquids, gels,syrups, slurries, suspensions and the like, for oral ingestion by asubject. In certain embodiments, pharmaceutical compositions for oraluse are obtained by mixing oligonucleotide and one or more solidexcipient. Suitable excipients include, but are not limited to, fillers,such as sugars, including lactose, sucrose, mannitol, or sorbitol;cellulose preparations such as, for example, maize starch, wheat starch,rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). In certain embodiments, such a mixture isoptionally ground and auxiliaries are optionally added. In certainembodiments, pharmaceutical compositions are formed to obtain tablets ordragee cores. In certain embodiments, disintegrating agents (e.g.,cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a saltthereof, such as sodium alginate) are added.

In certain embodiments, dragee cores are provided with coatings. Incertain such embodiments, concentrated sugar solutions may be used,which may optionally contain gum arabic, talc, polyvinyl pyrrolidone,carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquersolutions, and suitable organic solvents or solvent mixtures. Dyestuffsor pigments may be added to tablets or dragee coatings.

In certain embodiments, pharmaceutical compositions for oraladministration are push-fit capsules made of gelatin. Certain of suchpush-fit capsules comprise one or more pharmaceutical agents of thepresent invention in admixture with one or more filler such as lactose,binders such as starches, and/or lubricants such as talc or magnesiumstearate and, optionally, stabilizers. In certain embodiments,pharmaceutical compositions for oral administration are soft, sealedcapsules made of gelatin and a plasticizer, such as glycerol orsorbitol. In certain soft capsules, one or more pharmaceutical agents ofthe present invention are be dissolved or suspended in suitable liquids,such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added.

In certain embodiments, pharmaceutical compositions are prepared forbuccal administration. Certain of such pharmaceutical compositions aretablets or lozenges formulated in conventional manner.

In certain embodiments, a pharmaceutical composition is prepared foradministration by injection (e.g., intravenous, subcutaneous,intramuscular, etc.). In certain of such embodiments, a pharmaceuticalcomposition comprises a carrier and is formulated in aqueous solution,such as water or physiologically compatible buffers such as Hanks'ssolution, Ringer's solution, or physiological saline buffer. In certainembodiments, other ingredients are included (e.g., ingredients that aidin solubility or serve as preservatives). In certain embodiments,injectable suspensions are prepared using appropriate liquid carriers,suspending agents and the like. Certain pharmaceutical compositions forinjection are presented in unit dosage form, e.g., in ampoules or inmulti-dose containers. Certain pharmaceutical compositions for injectionare suspensions, solutions or emulsions in oily or aqueous vehicles, andmay contain formulatory agents such as suspending, stabilizing and/ordispersing agents. Certain solvents suitable for use in pharmaceuticalcompositions for injection include, but are not limited to, lipophilicsolvents and fatty oils, such as sesame oil, synthetic fatty acidesters, such as ethyl oleate or triglycerides, and liposomes. Aqueousinjection suspensions may contain substances that increase the viscosityof the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, such suspensions may also contain suitablestabilizers or agents that increase the solubility of the pharmaceuticalagents to allow for the preparation of highly concentrated solutions.

In certain embodiments, a pharmaceutical composition is prepared fortransmucosal administration. In certain of such embodiments penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

In certain embodiments, a pharmaceutical composition is prepared foradministration by inhalation. Certain of such pharmaceuticalcompositions for inhalation are prepared in the form of an aerosol sprayin a pressurized pack or a nebulizer. Certain of such pharmaceuticalcompositions comprise a propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In certain embodiments using a pressurized aerosol,the dosage unit may be determined with a valve that delivers a meteredamount. In certain embodiments, capsules and cartridges for use in aninhaler or insufflator may be formulated. Certain of such formulationscomprise a powder mixture of a pharmaceutical agent of the invention anda suitable powder base such as lactose or starch.

In certain embodiments, a pharmaceutical composition is prepared forrectal administration, such as a suppositories or retention enema.Certain of such pharmaceutical compositions comprise known ingredients,such as cocoa butter and/or other glycerides.

In certain embodiments, a pharmaceutical composition is prepared fortopical administration. Certain of such pharmaceutical compositionscomprise bland moisturizing bases, such as ointments or creams.Exemplary suitable ointment bases include, but are not limited to,petrolatum, petrolatum plus volatile silicones, lanolin and water in oilemulsions such as Eucerin™, available from Beiersdorf (Cincinnati,Ohio). Exemplary suitable cream bases include, but are not limited to,Nivea™ Cream, available from Beiersdorf (Cincinnati, Ohio), cold cream(USP), Purpose Cream™ available from Johnson & Johnson (New Brunswick,N.J.), hydrophilic ointment (USP) and Lubriderm™, available from Pfizer(Morris Plains, N.J.).

In certain embodiments, a pharmaceutical composition of the presentinvention comprises an oligonucleotide in a therapeutically effectiveamount. In certain embodiments, the therapeutically effective amount issufficient to prevent, alleviate or ameliorate symptoms of a disease orto prolong the survival of the subject being treated. Determination of atherapeutically effective amount is well within the capability of thoseskilled in the art.

In certain embodiments, one or more oligonucleotides of the presentinvention is formulated as a prodrug. In certain embodiments, upon invivo administration, a prodrug is chemically converted to thebiologically, pharmaceutically or therapeutically more active form ofthe oligonucleotide. In certain embodiments, prodrugs are useful becausethey are easier to administer than the corresponding active form. Forexample, in certain instances, a prodrug may be more bioavailable (e.g.,through oral administration) than is the corresponding active form. Incertain instances, a prodrug may have improved solubility compared tothe corresponding active form. In certain embodiments, prodrugs are lesswater soluble than the corresponding active form. In certain instances,such prodrugs possess superior transmittal across cell membranes, wherewater solubility is detrimental to mobility. In certain embodiments, aprodrug is an ester. In certain such embodiments, the ester ismetabolically hydrolyzed to carboxylic acid upon administration. Incertain instances the carboxylic acid containing compound is thecorresponding active form. In certain embodiments, a prodrug comprises ashort peptide (polyaminoacid) bound to an acid group. In certain of suchembodiments, the peptide is cleaved upon administration to form thecorresponding active form.

In certain embodiments, a prodrug is produced by modifying apharmaceutically active compound such that the active compound will beregenerated upon in vivo administration. The prodrug can be designed toalter the metabolic stability or the transport characteristics of adrug, to mask side effects or toxicity, to improve the flavor of a drugor to alter other characteristics or properties of a drug. By virtue ofknowledge of pharmacodynamic processes and drug metabolism in vivo,those of skill in this art, once a pharmaceutically active compound isknown, can design prodrugs of the compound (see, e.g., Nogrady (1985)Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392).

In certain embodiments, a pharmaceutical composition comprising one ormore pharmaceutical agents of the present invention is useful fortreating a conditions or disorders in a mammalian, and particularly in ahuman, subject. Suitable administration routes include, but are notlimited to, oral, rectal, transmucosal, intestinal, enteral, topical,suppository, through inhalation, intrathecal, intraventricular,intraperitoneal, intranasal, intraocular and parenteral (e.g.,intravenous, intramuscular, intramedullary, and subcutaneous). Incertain embodiments, pharmaceutical intrathecals are administered toachieve local rather than systemic exposures. For example,pharmaceutical compositions may be injected directly in the area ofdesired effect (e.g., in the renal or cardiac area).

In certain embodiments, a pharmaceutical composition of the presentinvention is administered in the form of a dosage unit (e.g., tablet,capsule, bolus, etc.). In certain embodiments, such pharmaceuticalcompositions comprise an oligonucleotide in a dose selected from 25 mg,30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg,130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg,175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg,220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg,265 mg, 270 mg, 270 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg,310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg, 350 mg,355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, 395 mg,400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg, 440 mg,445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg, 485 mg,490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg, 530 mg,535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg, 575 mg,580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg, 620 mg,625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg, 665 mg,670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg, 710 mg,715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg, 755 mg,760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg, and 800mg. In certain such embodiments, a pharmaceutical composition of thepresent invention comprises a dose of oligonucleotide selected from 25mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400mg, 500 mg, 600 mg, 700 mg, and 800 mg. In certain embodiments, apharmaceutical composition is comprises a dose of oligonucleotideselected from 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, and 400 mg.In certain embodiments the dose is administered at intervals rangingfrom more than once per day, once per day, once per week, twice perweek, three times per week, four times per week, five times per week, 6times per week, once per month to once per three months, for as long asneeded to sustain the desired effect.

In a further aspect, a pharmaceutical agent is sterile lyophilizedoligonucleotide that is reconstituted with a suitable diluent, e.g.,sterile water for injection. The reconstituted product is administeredas a subcutaneous injection or as an intravenous infusion after dilutioninto saline. The lyophilized drug product consists of theoligonucleotide which has been prepared in water for injection, adjustedto pH 7.0-9.0 with acid or base during preparation, and thenlyophilized. The lyophilized oligonucleotide may be 25-800 mg of theoligonucleotide. It is understood that this encompasses 25, 50, 75, 100,125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 425, 450, 475,500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, and 800 mgof lyophilized oligonucleotide. The lyophilized drug product may bepackaged in a 2 mL Type I, clear glass vial (ammonium sulfate-treated),stoppered with a bromobutyl rubber closure and sealed with an aluminumFLIP-OFF® overseal. In one embodiment, the lyophilized pharmaceuticalagent comprises ISIS 405879.

The compositions of the present invention may additionally contain otheradjunct components conventionally found in pharmaceutical compositions,at their art-established usage levels. Thus, for example, thecompositions may contain additional, compatible, pharmaceutically-activematerials such as, for example, antipruritics, astringents, localanesthetics or anti-inflammatory agents, or may contain additionalmaterials useful in physically formulating various dosage forms of thecompositions of the present invention, such as dyes, flavoring agents,preservatives, antioxidants, opacifiers, thickening agents andstabilizers. However, such materials, when added, should not undulyinterfere with the biological activities of the components of thecompositions of the present invention. The formulations can besterilized and, if desired, mixed with auxiliary agents, e.g.,lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, colorings, flavoringsand/or aromatic substances and the like which do not deleteriouslyinteract with the oligonucleotide(s) of the formulation.

Conjugated Antisense Compounds

Antisense compounds may be covalently linked to one or more moieties orconjugates which enhance the activity, visualization of cellulardistribution, or cellular uptake of the resulting antisenseoligonucleotides. Typical conjugate groups include cholesterol moietiesand lipid moieties. Additional conjugate groups include carbohydrates,phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone,acridine, fluoresceins, rhodamines, coumarins, and dyes.

Antisense compounds can also be modified to have one or more stabilizinggroups that are generally attached to one or both termini of antisensecompounds to enhance properties such as, for example, nucleasestability. Included in stabilizing groups are cap structures. Theseterminal modifications protect the antisense compound having terminalnucleic acid from exonuclease degradation, and can help in deliveryand/or localization within a cell. The cap can be present at the5′-terminus (5′-cap), or at the 3′-terminus (3′-cap), or can be presenton both termini. Cap structures are well known in the art and include,for example, inverted deoxy abasic caps. Further 3′ and 5′-stabilizinggroups that can be used to cap one or both ends of an antisense compoundto impart nuclease stability include those disclosed in WO 03/004602published on Jan. 16, 2003.

Cell Culture and Antisense Compounds Treatment

The effects of antisense compounds on the level, activity or expressionof PCSK9 nucleic acids can be tested in vitro in a variety of celltypes. Cell types used for such analyses are available from commercialvendors (e.g., American Type Culture Collection, Manassas, Va.; Zen-Bio,Inc., Research Triangle Park, N.C.; Clonetics Corporation, Walkersville,Md.) and cells are cultured according to the vendor's instructions usingcommercially available reagents (e.g., Invitrogen Life Technologies,Carlsbad, Calif.). Illustrative cell types include, but are not limitedto, HepG2 cells, Hep3B cells, and primary hepatocytes.

In Vitro Testing of Antisense Oligonucleotides

Described herein are methods for treatment of cells with antisenseoligonucleotides, which can be modified appropriately for treatment withother antisense compounds.

In general, cells are treated with antisense oligonucleotides when theycells reach approximately 60-80% confluency in culture.

One reagent commonly used to introduce antisense oligonucleotides intocultured cells includes the cationic lipid transfection reagentLIPOFECTIN® (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotidesare mixed with LIPOFECTIN® in OPTI-MEM® 1 (Invitrogen, Carlsbad, Calif.)to achieve the desired final concentration of antisense oligonucleotideand a LIPOFECTIN® concentration that typically ranges 2 to 12 ug/mL per100 nM antisense oligonucleotide.

Another reagent used to introduce antisense oligonucleotides intocultured cells includes LIPOFECTAMINE® (Invitrogen, Carlsbad, Calif.).Antisense oligonucleotide is mixed with LIPOFECTAMINE® in OPTI-MEM® 1reduced serum medium (Invitrogen, Carlsbad, Calif.) to achieve thedesired concentration of antisense oligonucleotide and a LIPOFECTAMINE®concentration that typically ranges 2 to 12 ug/mL per 100 nM antisenseoligonucleotide.

Cells are treated with antisense oligonucleotides by routine methods.Cells are typically harvested 16-24 hours after antisenseoligonucleotide treatment, at which time RNA or protein levels of targetnucleic acids are measured by methods known in the art and describedherein. In general, when treatments are performed in multiplereplicates, the data are presented as the average of the replicatetreatments.

The concentration of anti sense oligonucleotide used varies from cellline to cell line. Methods to determine the optimal antisenseoligonucleotide concentration for a particular cell line are well knownin the art. Antisense oligonucleotides are typically used atconcentrations ranging from 1 nM to 300 nM.

RNA Isolation

RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA.Methods of RNA isolation are well known in the art. RNA is preparedusing methods well known in the art, for example, using the TRIZOL®Reagent (Invitrogen, Carlsbad, Calif.) according to the manufacturer'srecommended protocols.

Analysis of Inhibition of Target Levels or Expression

Inhibition of levels or expression of a PCSK9 nucleic acid can beassayed in a variety of ways known in the art. For example, targetnucleic acid levels can be quantitated by, e.g., Northern blot analysis,competitive polymerase chain reaction (PCR), or quantitative real-timePCR. RNA analysis can be performed on total cellular RNA orpoly(A)+mRNA. Methods of RNA isolation are well known in the art.Northern blot analysis is also routine in the art. Quantitativereal-time PCR can be conveniently accomplished using the commerciallyavailable ABI PRISM® 7600, 7700, or 7900 Sequence Detection System,available from PE-Applied Biosystems, Foster City, Calif. and usedaccording to manufacturer's instructions.

Quantitative Real-Time PCR Analysis of Target RNA Levels

Quantitation of target RNA levels may be accomplished by quantitativereal-time PCR using the ABI PRISM® 7600, 7700, or 7900 SequenceDetection System (PE-Applied Biosystems, Foster City, Calif.) accordingto manufacturer's instructions. Methods of quantitative real-time PCRare well known in the art.

Prior to real-time PCR, the isolated RNA is subjected to a reversetranscriptase (RT) reaction, which produces complementary DNA (cDNA)that is then used as the substrate for the real-time PCR amplification.The RT and real-time PCR reactions are performed sequentially in thesame sample well. RT and real-time PCR reagents are obtained fromInvitrogen (Carlsbad, Calif.). RT, real-time-PCR reactions are carriedout by methods well known to those skilled in the art.

Gene (or RNA) target quantities obtained by real time PCR are normalizedusing either the expression level of a gene whose expression isconstant, such as GAPDH, or by quantifying total RNA using RIBOGREEN®(Invitrogen, Inc. Carlsbad, Calif.). GAPDH expression is quantified byreal time PCR, by being run simultaneously with the target,multiplexing, or separately. Total RNA is quantified using RIBOGREEN®RNA quantification reagent (Invitrogen, Inc. Carlsbad, Calif.). Methodsof RNA quantification by RIBOGREEN® are taught in Jones, L. J., et al,(Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR® 4000instrument (PE Applied Biosystems) is used to measure RIBOGREEN®fluorescence.

Probes and primers are designed to hybridize to a PCSK9 nucleic acid.Methods for designing real-time PCR probes and primers are well known inthe art, and may include the use of software such as PRIMER EXPRESS®Software (Applied Biosystems, Foster City, Calif.).

Analysis of Protein Levels

Antisense inhibition of PCSK9 nucleic acids can be assessed by measuringPCSK9 protein levels. Protein levels of PCSK9 can be evaluated orquantitated in a variety of ways well known in the art, such asimmunoprecipitation, Western blot analysis (immunoblotting),enzyme-linked immunosorbent assay (ELISA), quantitative protein assays,protein activity assays (for example, caspase activity assays),immunohistochemistry, immunocytochemistry or fluorescence-activated cellsorting (FACS). Antibodies directed to a target can be identified andobtained from a variety of sources, such as the MSRS catalog ofantibodies (Aerie Corporation, Birmingham, Mich.), or can be preparedvia conventional monoclonal or polyclonal antibody generation methodswell known in the art. Antibodies useful for the detection of human andrat PCSK9 are commercially available.

Serum Lipid Analysis

Plasma concentrations of total cholesterol, LDL-C, HDL-C, freecholesterol, triglycerides, glucose, ketones, transaminases, andphospholipids can be measured using an automated clinical chemistryanalyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein andcholesterol profiling can be performed as described by Crooke et al.,Lipid Res. (2005) 46: 872-884, using a Beckman System Gold 126 HPLCsystem, a 507e refrigerated autosampler, a 126 photodiode array detector(Beckman Instruments, Fullerton, Calif.), and a Superose 6 HR 10/30column (Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions are measuredat a wavelength of 505 nm and validated with a cholesterol calibrationkit. (Sigma).

Liver Triglyceride Analysis

Liver triglyceride content can be measured according to routineexperimental procedures, for example, per procedures described by Desaiet al., Diabetes (2001) 50: 2287-2295. Liver triglyceride content may bemeasured after antisense inhibition of PCSK9 for 6 weeks.

In Vivo Testing of Antisense Compounds

Antisense compounds, for example, antisense oligonucleotides, are testedin animals to assess their ability to inhibit expression of PCSK9 andproduce phenotypic changes, such as decreases in LDL-C. Testing may beperformed in normal animals, or in experimental disease models. Foradministration to animals, antisense oligonucleotides are formulated ina pharmaceutically acceptable diluent, such as phosphate-bufferedsaline. Administration includes parenteral routes of administration,such as intraperitoneal, intravenous, and subcutaneous. Calculation ofantisense oligonucleotide dosage and dosing frequency is within theabilities of those skilled in the art, and depends upon factors such asroute of administration and animal body weight. Following a period oftreatment with antisense oligonucleotides, RNA is isolated from varioustissues and changes in PCSK9 nucleic acid expression are measured.Changes in PCSK9 protein levels may also be measured.

Nonlimiting Disclosure and Incorporation by Reference

The following examples serve only to illustrate the methods, antisenseoligonucleotides, and compositions provided and are not intended tolimit the same. Each of the references, GENBANK® accession numbers, andthe like recited in the present application is incorporated herein byreference in its entirety.

EXAMPLES Example 1: Antisense Inhibition of Human PCSK9: A549 Cells

Antisense oligonucleotides targeted to a PCSK9 nucleic acid were testedfor their effects on PCSK9 mRNA in vitro. Cultured A549 cells at adensity of 4000 cells per well in a 96-well plate were treated with 60nM of antisense oligonucleotide. After a treatment period ofapproximately 24 hours, RNA was isolated from the cells and PCSK9 mRNAlevels were measured by quantitative real-time PCR, as described herein.PCSK9 mRNA levels were adjusted according to total RNA content asmeasured by RIBOGREEN®. Results are presented as percent inhibition ofPCSK9, relative to untreated control cells. Antisense oligonucleotidesthat exhibited at least 30% inhibition of PCSK9 expression are shown inTable 16.

The motif column indicates the wing-gap-wing motif of each antisenseoligonucleotide. Antisense oligonucleotides were designed as 5-10-5gapmers, or alternatively as 3-14-3 gapmers, where the gap segmentcomprises 2′-deoxynucleotides and each wing segment comprises 2′-MOEnucleotides. As illustrated in Table 16, a single nucleobase sequencemay be represented by a 5-10-5 motif as well as a 3-14-3 motif. “5′target site” indicates the 5′-most nucleotide which the antisenseoligonucleotide is targeted on the indicated GENBANK Accession No.

TABLE 16 Antisense inhibition of PCSK9 in human cells (Cell type A549)SEQ ID 5′ Target Target Nucleic % IsisNo NO Site Acid Sequence MotifInhibition 395150 5 242 NM_174936.2 GAGGAGACCTAGAGGCCGTG 5-10-5 57395151 6 300 NM_174936.2 AGGACCGCCTGGAGCTGACG 5-10-5 77 395152 7 410NM_174936.2 ACGCAAGGCTAGCACCAGCT 5-10-5 81 395153 9 480 NM_174936.2CCTTGGCGCAGCGGTGGAAG 5-10-5 51 395154 10 561 NM_174936.2GGCGGGCAGTGCGCTCTGAC 5-10-5 40 395155 11 600 NM_174936.2TGGTGAGGTATCCCCGGCGG 5-10-5 79 395156 14 620 NM_174936.2ATGGAAGACATGCAGGATCT 5-10-5 75 395157 15 646 NM_174936.2TTCACCAGGAAGCCAGGAAG 5-10-5 72 395158 17 705 NM_174936.2CCTCGATGTAGTCGACATGG 5-10-5 61 395159 18 785 NM_174936.2GTATTCATCCGCCCGGTACC 5-10-5 35 395160 19 835 NM_174936.2CTGGTGTCTAGGAGATACAC 5-10-5 47 395161 21 860 NM_174936.2GATTTCCCGGTGGTCACTCT 5-10-5 70 395162 24 890 NM_174936.2CTCGAAGTCGGTGACCATGA 5-10-5 52 395163 25 923 NM_174936.2GTGGAAGCGGGTCCCGTCCT 5-10-5 73 395164 26 970 NM_174936.2ACCCCTGCCAGGTGGGTGCC 5-10-5 69 395165 28 1004 NM_174936.2ACCCTTGGCCACGCCGGCAT 5-10-5 81 395166 29 1040 NM_174936.2TTGGCAGTTGAGCACGCGCA 5-10-5 65 395167 31 1077 NM_174936.2CCAGGCCTATGAGGGTGCCG 5-10-5 34 395168 32 1098 NM_174936.2GCTGGCTTTTCCGAATAAAC 5-10-5 69 395169 33 1210 NM_174936.2GTGACCAGCACGACCCCAGC 5-10-5 61 395171 34 1326 NM_174936.2CCAAAGTCCCCAGGGTCACC 5-10-5 38 395173 36 1340 NM_174936.2GCCAAAGTTGGTCCCCAAAG 5-10-5 64 395174 38 1361 NM_174936.2GGCAAAGAGGTCCACACAGC 5-10-5 72 395175 39 1389 NM_174936.2TGGAGGCACCAATGATGTCC 5-10-5 41 395177 45 1534 NM_174936.2TTGGCAGAGAAGTGGATCAG 5-10-5 41 395178 50 1569 NM_174936.2GGTCCTCAGGGAACCAGGCC 5-10-5 67 395181 52 1640 NM_174936.2AAACAGCTGCCAACCTGCCC 5-10-5 37 395182 54 1675 NM_174936.2GTAGGCCCCGAGTGTGCTGA 5-10-5 51 395183 57 1812 NM_174936.2CGTTGTGGGCCCGGCAGACC 5-10-5 74 395184 58 1858 NM_174936.2AGCAGGCAGCACCTGGCAAT 5-10-5 61 395185 59 1920 NM_174936.2CACGGGTCCCCATGCTGGCC 5-10-5 73 395186 60 2100 NM_174936.2CTTTGCATTCCAGACCTGGG 5-10-5 83 395187 62 2310 NM_174936.2GGCAGCAGATGGCAACGGCT 5-10-5 80 395189 64 2504 NM_174936.2TCAAGGGCCAGGCCAGCAGC 5-10-5 65 395190 66 2597 NM_174936.2ATGCCCCACAGTGAGGGAGG 5-10-5 60 395191 67 2606 NM_174936.2AATGGTGAAATGCCCCACAG 5-10-5 76 395193 68 2750 NM_174936.2CATGGGAAGAATCCTGCCTC 5-10-5 48 395194 69 2832 NM_174936.2GGAGATGAGGGCCATCAGCA 5-10-5 64 395195 70 2900 NM_174936.2TAGATGCCATCCAGAAAGCT 5-10-5 44 395196 72 2983 NM_174936.2GGCATAGAGCAGAGTAAAGG 5-10-5 39 395198 75 3437 NM_174936.2GCTCAAGGAGGGACAGTTGT 5-10-5 73 395199 76 3472 NM_174936.2AAAGATAAATGTCTGCTTGC 5-10-5 83 395200 78 3543 NM_174936.2TCTTCAAGTTACAAAAGCAA 5-10-5 40 395202 129 13816 NT_032977.8GTGCCATCTGAACAGCACCT 5-10-5 36 395203 110 13926 NT_032977.8CCTGGAACCCCTGCAGCCAG 5-10-5 64 395204 152 13977 NT_032977.8TTCAGGCAGGTTGCTGCTAG 5-10-5 55 395206 136 14122 NT_032977.8TAGGAGAAAGTAGGGAGAGC 5-10-5 45 395207 132 14179 NT_032977.8TAAAAGCTGCAAGAGACTCA 5-10-5 49 395208 139 14267 NT_032977.8TCAGAGAAAACAGTCACCGA 5-10-5 56 395209 142 14404 NT_032977.8TCATTTTAGAGACAGGAAGC 5-10-5 73 395210 113 14441 NT_032977.8GAATAACAGTGATGTCTGGC 5-10-5 82 395211 138 14494 NT_032977.8TCACAGCTCACCGAGTCTGC 5-10-5 60 395212 98 14524 NT_032977.8AGTGTAAAATAAAGCCCCTA 5-10-5 50 395213 96 14601 NT_032977.8AGGACCCAAGTCATCCTGCT 5-10-5 57 395214 124 14631 NT_032977.8GGCCATCAGCTGGCAATGCT 5-10-5 50 395215 133 14675 NT_032977.8TAGACAAGGAAAGGGAGGCC 5-10-5 60 395216 103 14681 NT_032977.8ATTTCATAGACAAGGAAAGG 5-10-5 51 395218 146 1315 AK124635.1TGACATTTGTGGGAGAGGAG 5-10-5 35 395220 104 2085 AK124635.1CACAGTCCTGCAAAACAGCT 5-10-5 54 395221 102 5590 NT_032977.8ATGTGCAGAGATCAATCACA 5-10-5 69 395222 127 10633 NT_032977.8GGTGGTAATTTGTCACAGCA 5-10-5 64 395223 84 11308 NT_032977.8AAGGTCACACAGTTAAGAGT 5-10-5 55 395226 147 24858 NT_032977.8TGAGTTCATTTAAGAGTGGA 5-10-5 42 399793 8 417 NM_174936.2CCTCGGAACGCAAGGCTAGC 5-10-5 71 399794 12 606 NM_174936.2GGATCTTGGTGAGGTATCCC 5-10-5 42 399795 13 615 NM_174936.2AGACATGCAGGATCTTGGTG 5-10-5 54 399796 16 651 NM_174936.2TCATCTTCACCAGGAAGCCA 5-10-5 50 399797 20 840 NM_174936.2GTATGCTGGTGTCTAGGAGA 5-10-5 38 399798 22 866 NM_174936.2GCCCTCGATTTCCCGGTGGT 5-10-5 54 399799 23 880 NM_174936.2GTGACCATGACCCTGCCCTC 5-10-5 75 399800 27 975 NM_174936.2TGACCACCCCTGCCAGGTGG 5-10-5 67 399801 30 1045 NM_174936.2TTCCCTTGGCAGTTGAGCAC 5-10-5 79 399802 35 1335 NM_174936.2AGTTGGTCCCCAAAGTCCCC 5-10-5 71 399803 37 1352 NM_174936.2GTCCACACAGCGGCCAAAGT 5-10-5 63 399804 40 1400 NM_174936.2GCTGCAGTCGCTGGAGGCAC 5-10-5 46 399805 41 1411 NM_174936.2ACAAAGCAGGTGCTGCAGTC 5-10-5 46 399806 42 1470 NM_174936.2GCATCATGGCTGCAATGCCA 5-10-5 85 399807 43 1478 NM_174936.2GGCAGACAGCATCATGGCTG 5-10-5 71 399808 44 1526 NM_174936.2GAAGTGGATCAGTCTCTGCC 5-10-5 38 399809 46 1539 NM_174936.2CATCTTTGGCAGAGAAGTGG 5-10-5 53 399810 47 1545 NM_174936.2TGATGACATCTTTGGCAGAG 5-10-5 63 399811 48 1552 NM_174936.2GCCTCATTGATGACATCTTT 5-10-5 61 399812 49 1564 NM_174936.2TCAGGGAACCAGGCCTCATT 5-10-5 46 399813 51 1583 NM_174936.2GGTCAGTACCCGCTGGTCCT 5-10-5 72 399814 53 1645 NM_174936.2CTGCAAAACAGCTGCCAACC 5-10-5 88 399815 55 1740 NM_174936.2AGAAACTGGAGCAGCTCAGC 5-10-5 38 399816 56 1746 NM_174936.2TCCTGGAGAAACTGGAGCAG 5-10-5 51 399817 61 2105 NM_174936.2CTTGACTTTGCATTCCAGAC 5-10-5 84 399818 63 2415 NM_174936.2AACCATTTTAAAGCTCAGCC 5-10-5 68 399819 65 2509 NM_174936.2CCCACTCAAGGGCCAGGCCA 5-10-5 87 399821 71 2906 NM_174936.2TCTGGCTAGATGCCATCCAG 5-10-5 83 399822 73 2988 NM_174936.2AGCCTGGCATAGAGCAGAGT 5-10-5 70 399824 74 3233 NM_174936.2AAGTAAGAAGAGGCTTGGCT 5-10-5 43 399825 77 3477 NM_174936.2ACCCAAAAGATAAATGTCTG 5-10-5 61 399827 100 13746 NT_032977.8ATCTCAGGACAGGTGAGCAA 5-10-5 81 399828 116 13760 NT_032977.8GAGTAGAGATTCTCATCTCA 5-10-5 73 399829 117 13828 NT_032977.8GAGTCTTCTGAAGTGCCATC 5-10-5 65 399831 83 13986 NT_032977.8AAGGAAGACTTCAGGCAGGT 5-10-5 52 399833 92 14397 NT_032977.8AGAGACAGGAAGCTGCAGCT 5-10-5 54 399834 82 14670 NT_032977.8AAGGAAAGGGAGGCCTAGAG 5-10-5 44 399836 130 2095 AK124635.1GTGCTGACCACACAGTCCTG 5-10-5 96 399837 107 3056 NT_032977.8CCCACTATAATGGCAAGCCC 5-10-5 83 399838 80 4306 NT_032977.8AACCCAGTTCTAATGCACCT 5-10-5 81 399839 106 5140 NT_032977.8CCAGTCAGAGTAGAACAGAG 5-10-5 68 399840 121 5599 NT_032977.8GGAGCCTACATGTGCAGAGA 5-10-5 46 399841 94 5667 NT_032977.8AGCATGGCACCAGCATCTGC 5-10-5 59 399842 108 6652 NT_032977.8CCCAGCCCTATCAGGAAGTG 5-10-5 67 399843 144 7099 NT_032977.8TGACATCCAGGAGGGAGGAG 5-10-5 33 399844 91 7556 NT_032977.8AGACTGATGGAAGGCATTGA 5-10-5 56 399845 131 7565 NT_032977.8GTGTTGAGCAGACTGATGGA 5-10-5 48 399846 145 8836 NT_032977.8TGACATCTTGTCTGGGAGCC 5-10-5 58 399847 90 8948 NT_032977.8AGACTAGGAGCCTGAGTTTT 5-10-5 38 399849 148 10252 NT_032977.8TGGCAGCAACTCAGACATAT 5-10-5 74 399851 88 12715 NT_032977.8ACTGGATACATTGGCAGACA 5-10-5 62 399852 111 12928 NT_032977.8CTAGAGGAACCACTAGATAT 5-10-5 66 399854 97 16134 NT_032977.8AGTCAAGCTGCTGCCCAGAG 5-10-5 82 399855 120 16668 NT_032977.8GCTAGTTATTAAGCACCTGC 5-10-5 71 399856 150 17267 NT_032977.8TGTGAGCTCTGGCCCAGTGG 5-10-5 64 399857 115 18377 NT_032977.8GAGTAAGGCAGGTTACTCTC 5-10-5 79 399858 134 18408 NT_032977.8TAGATGTGACTAACATTTAA 5-10-5 57 399859 105 19203 NT_032977.8CACATTAGCCTTGCTCAAGT 5-10-5 75 399860 151 19913 NT_032977.8TGTGATGACCTGGAAAGGTG 5-10-5 36 399862 109 20188 NT_032977.8CCCCTGCACAGAGCCTGGCA 5-10-5 59 399863 141 20624 NT_032977.8TCATGGCTGCAATGCCTGGT 5-10-5 46 399864 93 20995 NT_032977.8AGAGAGGAGGGCTTAAAGAA 5-10-5 30 399865 95 21082 NT_032977.8AGCTGCCAACCTGCAAAAAG 5-10-5 75 399866 143 21481 NT_032977.8TGAAAATCCATCCAGCACTG 5-10-5 74 399867 89 21589 NT_032977.8AGAACCATGGAGCACCTGAG 5-10-5 84 399868 123 21696 NT_032977.8GGCACTGCCCTTCCACCAAA 5-10-5 53 399869 118 24907 NT_032977.8GCACCATCCAGACCAGAATC 5-10-5 49 399870 114 25413 NT_032977.8GAGAGGTTCAGATCCAGGCC 5-10-5 66 399871 4 135 NM_174936.2GCGCGGAATCCTGGCTGGGA 3-14-3 40 399872 5 242 NM_174936.2GAGGAGACCTAGAGGCCGTG 3-14-3 64 399873 6 300 NM_174936.2AGGACCGCCTGGAGCTGACG 3-14-3 65 399874 7 410 NM_174936.2ACGCAAGGCTAGCACCAGCT 3-14-3 80 399875 9 480 NM_174936.2CCTTGGCGCAGCGGTGGAAG 3-14-3 67 399876 10 561 NM_174936.2GGCGGGCAGTGCGCTCTGAC 3-14-3 62 399877 11 600 NM_174936.2TGGTGAGGTATCCCCGGCGG 3-14-3 73 399878 14 620 NM_174936.2ATGGAAGACATGCAGGATCT 3-14-3 54 399879 15 646 NM_174936.2TTCACCAGGAAGCCAGGAAG 3-14-3 75 399880 17 705 NM_174936.2CCTCGATGTAGTCGACATGG 3-14-3 80 399881 18 785 NM_174936.2GTATTCATCCGCCCGGTACC 3-14-3 63 399882 19 835 NM_174936.2CTGGTGTCTAGGAGATACAC 3-14-3 66 399883 21 860 NM_174936.2GATTTCCCGGTGGTCACTCT 3-14-3 59 399884 24 890 NM_174936.2CTCGAAGTCGGTGACCATGA 3-14-3 71 399885 25 923 NM_174936.2GTGGAAGCGGGTCCCGTCCT 3-14-3 73 399886 26 970 NM_174936.2ACCCCTGCCAGGTGGGTGCC 3-14-3 76 399887 28 1004 NM_174936.2ACCCTTGGCCACGCCGGCAT 3-14-3 54 399888 29 1040 NM_174936.2TTGGCAGTTGAGCACGCGCA 3-14-3 75 399889 31 1077 NM_174936.2CCAGGCCTATGAGGGTGCCG 3-14-3 84 399890 32 1098 NM_174936.2GCTGGCTTTTCCGAATAAAC 3-14-3 73 399891 33 1210 NM_174936.2GTGACCAGCACGACCCCAGC 3-14-3 99 399892 149 1297 NM_174936.2TGGGCATTGGTGGCCCCAAC 3-14-3 61 399893 34 1326 NM_174936.2CCAAAGTCCCCAGGGTCACC 3-14-3 54 399894 128 1330 NM_174936.2GTCCCCAAAGTCCCCAGGGT 3-14-3 80 399895 36 1340 NM_174936.2GCCAAAGTTGGTCCCCAAAG 3-14-3 82 399896 38 1361 NM_174936.2GGCAAAGAGGTCCACACAGC 3-14-3 83 399897 39 1389 NM_174936.2TGGAGGCACCAATGATGTCC 3-14-3 70 399898 101 1465 NM_174936.2ATGGCTGCAATGCCAGCCAC 3-14-3 37 399899 45 1534 NM_174936.2TTGGCAGAGAAGTGGATCAG 3-14-3 64 399900 50 1569 NM_174936.2GGTCCTCAGGGAACCAGGCC 3-14-3 82 399901 87 1576 NM_174936.2ACCCGCTGGTCCTCAGGGAA 3-14-3 76 399902 119 1605 NM_174936.2GCAGGGCGGCCACCAGGTTG 3-14-3 39 399903 52 1640 NM_174936.2AAACAGCTGCCAACCTGCCC 3-14-3 73 399904 54 1675 NM_174936.2GTAGGCCCCGAGTGTGCTGA 3-14-3 81 399905 57 1812 NM_174936.2CGTTGTGGGCCCGGCAGACC 3-14-3 80 399906 58 1858 NM_174936.2AGCAGGCAGCACCTGGCAAT 3-14-3 92 399907 59 1920 NM_174936.2CACGGGTCCCCATGCTGGCC 3-14-3 95 399908 60 2100 NM_174936.2CTTTGCATTCCAGACCTGGG 3-14-3 84 399909 62 2310 NM_174936.2GGCAGCAGATGGCAACGGCT 3-14-3 73 399910 154 2410 NM_174936.2TTTTAAAGCTCAGCCCCAGC 3-14-3 57 399911 64 2504 NM_174936.2TCAAGGGCCAGGCCAGCAGC 3-14-3 82 399912 66 2597 NM_174936.2ATGCCCCACAGTGAGGGAGG 3-14-3 68 399913 67 2606 NM_174936.2AATGGTGAAATGCCCCACAG 3-14-3 92 399914 153 2649 NM_174936.2TTGGGAGCAGCTGGCAGCAC 3-14-3 59 399915 68 2750 NM_174936.2CATGGGAAGAATCCTGCCTC 3-14-3 78 399916 69 2832 NM_174936.2GGAGATGAGGGCCATCAGCA 3-14-3 71 399917 70 2900 NM_174936.2TAGATGCCATCCAGAAAGCT 3-14-3 81 399918 72 2983 NM_174936.2GGCATAGAGCAGAGTAAAGG 3-14-3 82 399919 112 3227 NM_174936.2GAAGAGGCTTGGCTTCAGAG 3-14-3 57 399920 75 3437 NM_174936.2GCTCAAGGAGGGACAGTTGT 3-14-3 84 399921 76 3472 NM_174936.2AAAGATAAATGTCTGCTTGC 3-14-3 78 399922 78 3543 NM_174936.2TCTTCAAGTTACAAAAGCAA 3-14-3 61 399923 85 13681 NT_032977.8ACAAATTCCCAGACTCAGCA 3-14-3 82 399924 129 13816 NT_032977.8GTGCCATCTGAACAGCACCT 3-14-3 84 399925 110 13926 NT_032977.8CCTGGAACCCCTGCAGCCAG 3-14-3 73 399926 152 13977 NT_032977.8TTCAGGCAGGTTGCTGCTAG 3-14-3 47 399927 140 13998 NT_032977.8TCAGCCAGGCCAAAGGAAGA 3-14-3 78 399928 136 14122 NT_032977.8TAGGAGAAAGTAGGGAGAGC 3-14-3 47 399929 132 14179 NT_032977.8TAAAAGCTGCAAGAGACTCA 3-14-3 69 399930 139 14267 NT_032977.8TCAGAGAAAACAGTCACCGA 3-14-3 82 399931 142 14404 NT_032977.8TCATTTTAGAGACAGGAAGC 3-14-3 79 399932 113 14441 NT_032977.8GAATAACAGTGATGTCTGGC 3-14-3 83 399933 138 14494 NT_032977.8TCACAGCTCACCGAGTCTGC 3-14-3 78 399934 98 14524 NT_032977.8AGTGTAAAATAAAGCCCCTA 3-14-3 79 399935 96 14601 NT_032977.8AGGACCCAAGTCATCCTGCT 3-14-3 89 399936 124 14631 NT_032977.8GGCCATCAGCTGGCAATGCT 3-14-3 70 399937 133 14675 NT_032977.8TAGACAAGGAAAGGGAGGCC 3-14-3 78 399938 103 14681 NT_032977.8ATTTCATAGACAAGGAAAGG 3-14-3 67 399940 146 1315 AK124635.1TGACATTTGTGGGAGAGGAG 3-14-3 35 399942 104 2085 AK124635.1CACAGTCCTGCAAAACAGCT 3-14-3 75 399943 102 5590 NT_032977.8ATGTGCAGAGATCAATCACA 3-14-3 64 399944 127 10633 NT_032977.8GGTGGTAATTTGTCACAGCA 3-14-3 73 399945 84 11308 NT_032977.8AAGGTCACACAGTTAAGAGT 3-14-3 89 399947 126 22292 NT_032977.8GGTGCATAAGGAGAAAGAGA 3-14-3 70 399948 147 24858 NT_032977.8TGAGTTCATTTAAGAGTGGA 3-14-3 46 399949 8 417 NM_174936.2CCTCGGAACGCAAGGCTAGC 3-14-3 74 399950 12 606 NM_174936.2GGATCTTGGTGAGGTATCCC 3-14-3 68 399951 13 615 NM_174936.2AGACATGCAGGATCTTGGTG 3-14-3 67 399952 16 651 NM_174936.2TCATCTTCACCAGGAAGCCA 3-14-3 74 399953 20 840 NM_174936.2GTATGCTGGTGTCTAGGAGA 3-14-3 59 399954 22 866 NM_174936.2GCCCTCGATTTCCCGGTGGT 3-14-3 83 399955 23 880 NM_174936.2GTGACCATGACCCTGCCCTC 3-14-3 62 399956 27 975 NM_174936.2TGACCACCCCTGCCAGGTGG 3-14-3 78 399957 30 1045 NM_174936.2TTCCCTTGGCAGTTGAGCAC 3-14-3 82 399958 35 1335 NM_174936.2AGTTGGTCCCCAAAGTCCCC 3-14-3 78 399959 37 1352 NM_174936.2GTCCACACAGCGGCCAAAGT 3-14-3 89 399960 40 1400 NM_174936.2GCTGCAGTCGCTGGAGGCAC 3-14-3 64 399961 41 1411 NM_174936.2ACAAAGCAGGTGCTGCAGTC 3-14-3 63 399962 42 1470 NM_174936.2GCATCATGGCTGCAATGCCA 3-14-3 68 399963 43 1478 NM_174936.2GGCAGACAGCATCATGGCTG 3-14-3 70 399964 44 1526 NM_174936.2GAAGTGGATCAGTCTCTGCC 3-14-3 70 399965 46 1539 NM_174936.2CATCTTTGGCAGAGAAGTGG 3-14-3 49 399966 47 1545 NM_174936.2TGATGACATCTTTGGCAGAG 3-14-3 78 399967 48 1552 NM_174936.2GCCTCATTGATGACATCTTT 3-14-3 82 399968 49 1564 NM_174936.2TCAGGGAACCAGGCCTCATT 3-14-3 71 399969 51 1583 NM_174936.2GGTCAGTACCCGCTGGTCCT 3-14-3 82 399970 53 1645 NM_174936.2CTGCAAAACAGCTGCCAACC 3-14-3 74 399971 55 1740 NM_174936.2AGAAACTGGAGCAGCTCAGC 3-14-3 48 399972 56 1746 NM_174936.2TCCTGGAGAAACTGGAGCAG 3-14-3 63 399973 61 2105 NM_174936.2CTTGACTTTGCATTCCAGAC 3-14-3 74 399974 63 2415 NM_174936.2AACCATTTTAAAGCTCAGCC 3-14-3 71 399975 65 2509 NM_174936.2CCCACTCAAGGGCCAGGCCA 3-14-3 91 399976 122 2582 NM_174936.2GGAGGGAGCTTCCTGGCACC 3-14-3 67 399977 71 2906 NM_174936.2TCTGGCTAGATGCCATCCAG 3-14-3 89 399978 73 2988 NM_174936.2AGCCTGGCATAGAGCAGAGT 3-14-3 80 399979 135 2994 NM_174936.2TAGCACAGCCTGGCATAGAG 3-14-3 83 399980 74 3233 NM_174936.2AAGTAAGAAGAGGCTTGGCT 3-14-3 50 399981 77 3477 NM_174936.2ACCCAAAAGATAAATGTCTG 3-14-3 55 399982 99 3550 NM_174936.2ATAAATATCTTCAAGTTACA 3-14-3 36 399983 100 13746 NT_032977.8ATCTCAGGACAGGTGAGCAA 3-14-3 96 399984 116 13760 NT_032977.8GAGTAGAGATTCTCATCTCA 3-14-3 71 399985 117 13828 NT_032977.8GAGTCTTCTGAAGTGCCATC 3-14-3 76 399986 81 13903 NT_032977.8AAGCAGGGCCTCAGGTGGAA 3-14-3 68 399987 83 13986 NT_032977.8AAGGAAGACTTCAGGCAGGT 3-14-3 59 399988 137 14112 NT_032977.8TAGGGAGAGCTCACAGATGC 3-14-3 31 399989 92 14397 NT_032977.8AGAGACAGGAAGCTGCAGCT 3-14-3 65 399990 82 14670 NT_032977.8AAGGAAAGGGAGGCCTAGAG 3-14-3 59 399992 130 2095 AK124635.1GTGCTGACCACACAGTCCTG 3-14-3 94 399993 107 3056 NT_032977.8CCCACTATAATGGCAAGCCC 3-14-3 76 399994 80 4306 NT_032977.8AACCCAGTTCTAATGCACCT 3-14-3 82 399995 106 5140 NT_032977.8CCAGTCAGAGTAGAACAGAG 3-14-3 74 399996 121 5599 NT_032977.8GGAGCCTACATGTGCAGAGA 3-14-3 65 399997 94 5667 NT_032977.8AGCATGGCACCAGCATCTGC 3-14-3 77 399998 108 6652 NT_032977.8CCCAGCCCTATCAGGAAGTG 3-14-3 78 399999 144 7099 NT_032977.8TGACATCCAGGAGGGAGGAG 3-14-3 43 400000 91 7556 NT_032977.8AGACTGATGGAAGGCATTGA 3-14-3 84 400001 131 7565 NT_032977.8GTGTTGAGCAGACTGATGGA 3-14-3 73 400002 145 8836 NT_032977.8TGACATCTTGTCTGGGAGCC 3-14-3 82 400003 90 8948 NT_032977.8AGACTAGGAGCCTGAGTTTT 3-14-3 48 400004 125 9099 NT_032977.8GGCCTGCAGAAGCCAGAGAG 3-14-3 45 400005 148 10252 NT_032977.8TGGCAGCAACTCAGACATAT 3-14-3 73 400006 79 11472 NT_032977.8AAATGCAGGGCTAAAATCAC 3-14-3 78 400007 88 12715 NT_032977.8ACTGGATACATTGGCAGACA 3-14-3 77 400008 111 12928 NT_032977.8CTAGAGGAACCACTAGATAT 3-14-3 78 400009 86 15471 NT_032977.8ACAGCATTCTTGGTTAGGAG 3-14-3 73 400010 97 16134 NT_032977.8AGTCAAGCTGCTGCCCAGAG 3-14-3 74 400011 120 16668 NT_032977.8GCTAGTTATTAAGCACCTGC 3-14-3 75 400012 150 17267 NT_032977.8TGTGAGCTCTGGCCCAGTGG 3-14-3 64 400013 115 18377 NT_032977.8GAGTAAGGCAGGTTACTCTC 3-14-3 88 400014 134 18408 NT_032977.8TAGATGTGACTAACATTTAA 3-14-3 58 400015 105 19203 NT_032977.8CACATTAGCCTTGCTCAAGT 3-14-3 89 400018 109 20188 NT_032977.8CCCCTGCACAGAGCCTGGCA 3-14-3 62 400019 141 20624 NT_032977.8TCATGGCTGCAATGCCTGGT 3-14-3 47 400020 93 20995 NT_032977.8AGAGAGGAGGGCTTAAAGAA 3-14-3 66 400021 95 21082 NT_032977.8AGCTGCCAACCTGCAAAAAG 3-14-3 61 400022 143 21481 NT_032977.8TGAAAATCCATCCAGCACTG 3-14-3 90 400023 89 21589 NT_032977.8AGAACCATGGAGCACCTGAG 3-14-3 83 400024 123 21696 NT_032977.8GGCACTGCCCTTCCACCAAA 3-14-3 79 400025 118 24907 NT_032977.8GCACCATCCAGACCAGAATC 3-14-3 78 400026 114 25413 NT_032977.8GAGAGGTTCAGATCCAGGCC 3-14-3 81

Antisense oligonucleotides with the following ISIS Nos exhibited atleast 80% inhibition of PCSK9 mRNA levels: 399806, 399814, 399817,399819, 399821, 399827, 399836, 399837, 399854, 399867, 399889, 399891,399906, 399907, 399908, 399913, 399920, 399924, 399935, 399945, 399959,399975, 399977, 399983, 399992, 400000, 400013, 400015, 400022, and400023. The target segments to which these antisense oligonucleotidesare targeted are active target segments. The target regions to whichthese antisense oligonucleotides are targeted are active target regions.

ISIS Nos 399814, 399819, 399836, 399891, 399906, 399907, 399913, 399935,399945, 399959, 399975, 399977, 399983, 399992, 400013, 400015, and400022 each exhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 399836, 399891, 399906, 399907, 399913, 399975, 399983, 399992,and 400022 each exhibited at least 90% inhibition of PCSK9 mRNA levels.

Example 2: Antisense Inhibition of Human PCSK9 in HepG2 Cells

Antisense oligonucleotides were tested for their ability to inhibit theexpression of PCSK9 mRNA in cultured HepG2 cells. HepG2 cells at adensity of 10000 cells per well in a 96-well plate were treated with1500 nM of antisense oligonucleotide. After a treatment period ofapproximately 24 hours, RNA was isolated from the cells and PCSK9 mRNAlevels were measured by quantitative real-time PCR, as described herein.PCSK9 mRNA levels were adjusted according to total RNA content asmeasured by RIBOGREEN®. Results are presented as percent inhibition ofPCSK9, relative to untreated control cells.

Isis Nos. 399819, 395149, 395150, 395151, 395152, 395153, 395154,395155, 395156, 395157, 395158, 395161, 395162, 395163, 395164, 395165,395166, 395167, 395168, 395172, 395173, 395174, 395178, 395179, 395182,395183, 395185, 395186, 395187, 395189, 395190, 395191, 395193, 395194,395195, 395198, 395199, 395203, 395207, 395210, 395211, 395213, 395214,395221, 395222, 399793, 399794, 399795, 399798, 399801, 399803, 399804,399806, 399807, 399812, 399813, 399814, 399816, 399817, 399820, 399821,399822, 399823, 399827, 399828, 399829, 399833, 399834, 399836, 399837,399841, 399844, 399846, 399848, 399849, 399851, 399852, 399854, 399855,399856, 399857, 399862, 399865, 399867, 399868, 399872, 399873, 399874,399875, 399876, 399877, 399879, 399880, 399883, 399884, 399885, 399886,399887, 399888, 399889, 399890, 399891, 399892, 399894, 399895, 399896,399900, 399901, 399902, 399904, 399905, 399906, 399907, 399908, 399909,399911, 399912, 399913, 399915, 399916, 399918, 399920, 399921, 399924,399925, 399929, 399931, 399935, 399936, 399937, 399943, 399944, 399945,399949, 399950, 399954, 399957, 399959, 399960, 399962, 399963, 399966,399967, 399968, 399969, 399970, 399972, 399973, 399975, 399976, 399977,399978, 399979, 399983, 399984, 399985, 399986, 399989, 399990, 399992,399996, 399997, 399998, 400000, 400001, 400002, 400004, 400005, 400007,400008, 400009, 400010, 400012, 400013, 400015, 400018, 400021, 400023,400024, 400025, and 400026 each inhibited PCSK9 by at least 70% in thisexperiment. The target segments to which these antisenseoligonucleotides are targeted are active target segments. The targetregions to which these antisense oligonucleotides are targeted areactive target regions.

Isis Nos 399819, 395150, 395152, 395153, 395154, 395155, 395156, 395158,395163, 395164, 395165, 395166, 395167, 395168, 395173, 395178, 395183,395185, 395186, 395187, 395189, 395190, 395193, 395194, 395199, 395203,395210, 395213, 395214, 399793, 399798, 399801, 399804, 399806, 399813,399816, 399820, 399821, 399822, 399836, 399849, 399856, 399857, 399862,399868, 399875, 399877, 399880, 399885, 399886, 399887, 399888, 399889,399890, 399894, 399896, 399900, 399901, 399904, 399905, 399906, 399907,399911, 399912, 399916, 399920, 399925, 399935, 399936, 399937, 399945,399954, 399957, 399959, 399960, 399962, 399968, 399969, 399973, 399976,399977, 399978, 399989, 399992, 400002, 400004, 400007, 400012, 400013,400023, and 400024 each inhibited PCSK9 by at least 80% in thisexperiment.

Isis Nos 399819, 395152, 395153, 395155, 395156, 395158, 395163, 395164,395165, 395166, 395168, 395178, 395183, 395185, 395187, 395189, 395190,395194, 395199, 395203, 395214, 399793, 399804, 399813, 399821, 399822,399836, 399856, 399868, 399877, 399887, 399888, 399890, 399900, 399904,399905, 399907, 399911, 399912, 399916, 399925, 399936, 399954, 399960,399962, 399969, 399976, 399977, 399978, 399989, 399992, 400012, 400013,400023, and 400024 each inhibited PCSK9 by at least 85% in thisexperiment.

Isis Nos 399819, 395165, 395183, 395185, 399868, 399900, 399907, 399911,399912, 399916, 399936, 399969, and 400024 each inhibited PCSK9 by atleast 90% in this experiment.

Example 3: Antisense Inhibition of Human PCSK9 (Hep3B Cells)

Antisense oligonucleotides targeted to a PCSK9 nucleic acid were testedfor their effects on PCSK9 mRNA in vitro. Cultured Hep3B cells at adensity of 4000 cells per well in a 96-well plate were treated with 75nM of antisense oligonucleotide. After a treatment period ofapproximately 24 hours, RNA was isolated from the cells and PCSK9 mRNAlevels were measured by quantitative real-time PCR, as described herein.PCSK9 mRNA levels were adjusted according to total RNA content asmeasured by RIBOGREEN®. Results are presented as percent inhibition ofPCSK9, relative to untreated control cells. Antisense oligonucleotidesthat exhibited at least 30% inhibition of PCSK9 expression are shown inTables 17, 18, and 19.

The motif column indicates the wing-gap-wing motif of each antisenseoligonucleotide. Antisense oligonucleotides were designed as 5-10-5gapmers, or alternatively as 3-14-3 gapmers, or alternatively as 2-13-5gapmers where the gap segment comprises 2′-deoxynucleotides and eachwing segment comprises 2′-MOE nucleotides. As illustrated in Table 17,18 and 19, a single nucleobase sequence may be represented by a 5-10-5motif as well as a 3-14-3 motif as well as a 2-13-5 motif. “5′ targetsite” indicates the 5′-most nucleotide which the antisenseoligonucleotide is targeted on the indicated GENBANK Accession No.

TABLE 17 Antisense inhibition of PCSK9 in human cells (Hep3B) targetingSEQ ID NO: 1 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % IDIsis No. NO: 1 NO: 1 Sequence 5′-3′ Motif inhibition NO 410742 294 313GCCTGGAGCTGACGGTGCCC 5-10-5 75.1 159 405999 298 317 GACCGCCTGGAGCTGACGGT5-10-5 70.3 160 395151 300 319 AGGACCGCCTGGAGCTGACG 5-10-5 57.3 6 405861406 425 AAGGCTAGCACCAGCTCCTC 5-10-5 90.5 162 405862 407 426CAAGGCTAGCACCAGCTCCT 5-10-5 92.3 163 405863 408 427 GCAAGGCTAGCACCAGCTCC5-10-5 82.6 164 405864 409 428 CGCAAGGCTAGCACCAGCTC 5-10-5 91.8 165395152 410 429 ACGCAAGGCTAGCACCAGCT 5-10-5 93.3 7 405865 411 430AACGCAAGGCTAGCACCAGC 5-10-5 84.6 166 405866 412 431 GAACGCAAGGCTAGCACCAG5-10-5 81.8 167 405867 413 432 GGAACGCAAGGCTAGCACCA 5-10-5 74.0 168405868 414 433 CGGAACGCAAGGCTAGCACC 5-10-5 76.8 169 399793 417 436CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 410743 421 440 TCCTCCTCGGAACGCAAGGC5-10-5 76.7 170 410744 446 465 GTGCTCGGGTGCTTCGGCCA 5-10-5 76.9 171410745 466 485 TGGAAGGTGGCTGTGGTTCC 5-10-5 41.7 172 395153 480 499CCTTGGCGCAGCGGTGGAAG 5-10-5 81.1 9 406000 482 501 ATCCTTGGCGCAGCGGTGGA5-10-5 79.1 173 406001 484 503 GGATCCTTGGCGCAGCGGTG 5-10-5 63.1 174406002 488 507 CCACGGATCCTTGGCGCAGC 5-10-5 80.8 175 410746 507 526CGTAGGTGCCAGGCAACCTC 5-10-5 44.8 176 410747 545 564 TGACTGCGAGAGGTGGGTCT5-10-5 NA 177 406003 555 574 CAGTGCGCTCTGACTGCGAG 5-10-5 84.2 178 406004557 576 GGCAGTGCGCTCTGACTGCG 5-10-5 56.4 179 406005 559 578CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 406006 562 581 CGGCGGGCAGTGCGCTCTGA5-10-5 71.0 181 410748 591 610 ATCCCCGGCGGGCAGCCTGG 5-10-5 56.0 182406007 595 614 AGGTATCCCCGGCGGGCAGC 5-10-5 76.3 183 410574 597 616TGAGGTATCCCCGGCGGGCA 3-14-3 69.1 184 410529 597 616 TGAGGTATCCCCGGCGGGCA5-10-5 77.4 184 410647 597 616 TGAGGTATCCCCGGCGGGCA 2-13-5 90.7 184410575 598 617 GTGAGGTATCCCCGGCGGGC 3-14-3 71.5 185 410648 598 617GTGAGGTATCCCCGGCGGGC 2-13-5 82.8 185 410530 598 617 GTGAGGTATCCCCGGCGGGC5-10-5 83.2 185 410649 599 618 GGTGAGGTATCCCCGGCGGG 2-13-5 53.5 186410576 599 618 GGTGAGGTATCCCCGGCGGG 3-14-3 56.2 186 410531 599 618GGTGAGGTATCCCCGGCGGG 5-10-5 63.6 186 410650 600 619 TGGTGAGGTATCCCCGGCGG2-13-5 86.1 11 399877 600 619 TGGTGAGGTATCCCCGGCGG 3-14-3 92.6 11 395155600 619 TGGTGAGGTATCCCCGGCGG 5-10-5 92.7 11 410577 601 620TTGGTGAGGTATCCCCGGCG 3-14-3 62.4 187 410532 601 620 TTGGTGAGGTATCCCCGGCG5-10-5 63.5 187 410651 601 620 TTGGTGAGGTATCCCCGGCG 2-13-5 75.5 187410652 602 621 CTTGGTGAGGTATCCCCGGC 2-13-5 81.2 188 410578 602 621CTTGGTGAGGTATCCCCGGC 3-14-3 83.3 188 406008 602 621 CTTGGTGAGGTATCCCCGGC5-10-5 95.2 188 410653 603 622 TCTTGGTGAGGTATCCCCGG 2-13-5 68.9 189410579 603 622 TCTTGGTGAGGTATCCCCGG 3-14-3 74.0 189 410533 603 622TCTTGGTGAGGTATCCCCGG 5-10-5 79.0 189 410580 604 623 ATCTTGGTGAGGTATCCCCG3-14-3 62.3 190 410654 604 623 ATCTTGGTGAGGTATCCCCG 2-13-5 65.2 190406009 604 623 ATCTTGGTGAGGTATCCCCG 5-10-5 90.7 190 410534 605 624GATCTTGGTGAGGTATCCCC 5-10-5 59.8 191 410581 605 624 GATCTTGGTGAGGTATCCCC3-14-3 72.9 191 410655 605 624 GATCTTGGTGAGGTATCCCC 2-13-5 77.4 191399794 606 625 GGATCTTGGTGAGGTATCCC 5-10-5 56.5 12 410656 606 625GGATCTTGGTGAGGTATCCC 2-13-5 68.0 12 399950 606 625 GGATCTTGGTGAGGTATCCC3-14-3 74.4 12 410535 607 626 AGGATCTTGGTGAGGTATCC 5-10-5 65.7 192410582 607 626 AGGATCTTGGTGAGGTATCC 3-14-3 69.5 192 410657 607 626AGGATCTTGGTGAGGTATCC 2-13-5 73.1 192 406010 609 628 GCAGGATCTTGGTGAGGTAT5-10-5 56.9 193 406011 611 630 ATGCAGGATCTTGGTGAGGT 5-10-5 70.7 194406012 613 632 ACATGCAGGATCTTGGTGAG 5-10-5 69.1 195 406013 617 636GAAGACATGCAGGATCTTGG 5-10-5 77.4 196 395156 620 639 ATGGAAGACATGCAGGATCT5-10-5 76.7 14 410749 628 647 AGAAGGCCATGGAAGACATG 5-10-5 59.4 197410750 638 657 GAAGCCAGGAAGAAGGCCAT 5-10-5 57.5 198 399879 646 665TTCACCAGGAAGCCAGGAAG 3-14-3 91.9 15 406014 648 667 TCTTCACCAGGAAGCCAGGA5-10-5 94.0 199 406015 653 672 ACTCATCTTCACCAGGAAGC 5-10-5 78.8 200406016 655 674 CCACTCATCTTCACCAGGAA 5-10-5 87.2 201 410730 657 676CGCCACTCATCTTCACCAGG 5-10-5 85.4 202 406017 659 678 GTCGCCACTCATCTTCACCA5-10-5 76.3 203 406018 661 680 AGGTCGCCACTCATCTTCAC 5-10-5 63.2 204406019 663 682 GCAGGTCGCCACTCATCTTC 5-10-5 68.1 205 406020 665 684CAGCAGGTCGCCACTCATCT 5-10-5 83.7 206 410731 667 686 TCCAGCAGGTCGCCACTCAT5-10-5 72.9 207 410751 685 704 GGCAACTTCAAGGCCAGCTC 5-10-5 84.6 208395158 705 724 CCTCGATGTAGTCGACATGG 5-10-5 93.9 17 410752 724 743GCAAAGACAGAGGAGTCCTC 5-10-5 76.3 209 406021 782 801 TTCATCCGCCCGGTACCGTG5-10-5 79.6 210 406022 784 803 TATTCATCCGCCCGGTACCG 5-10-5 78.7 211406023 787 806 TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 406024 789 808GCTGGTATTCATCCGCCCGG 5-10-5 69.7 213 410732 791 810 GGGCTGGTATTCATCCGCCC5-10-5 30.4 214 410753 821 840 ATACACCTCCACCAGGCTGC 5-10-5 72.8 215406025 832 851 GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 837 856TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 862 881 TCGATTTCCCGGTGGTCACT5-10-5 87.6 218 405870 863 882 CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219405871 864 883 CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 865 884CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 866 885 GCCCTCGATTTCCCGGTGGT5-10-5 84.3 22 399954 866 885 GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873867 886 TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 868 887CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 869 888 CCTGCCCTCGATTTCCCGGT5-10-5 93.5 224 405876 870 889 CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225406027 874 893 ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 876 895CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 878 897 GACCATGACCCTGCCCTCGA5-10-5 91.4 228 406030 882 901 CGGTGACCATGACCCTGCCC 5-10-5 95.6 229406031 884 903 GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 886 905AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 888 907 CGAAGTCGGTGACCATGACC5-10-5 88.5 232 410754 898 917 GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233395163 923 942 GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410755 933 952TGGCCTGTCTGTGGAAGCGG 5-10-5 NA 234 410756 960 979 GGTGGGTGCCATGACTGTCA5-10-5 67.1 235 406033 967 986 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237406034 972 991 CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 977 996GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 985 1004TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 989 1008GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 992 1011GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 997 1016GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 997 1016GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 997 1016GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 998 1017GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 998 1017GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 998 1017GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 999 1018TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 999 1018TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 999 1018TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 1000 1019TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 1000 1019TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 1000 1019TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 1001 1020CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 1001 1020CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 1001 1020CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 1002 1021CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 1002 1021CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 1002 1021CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 1003 1022CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 1003 1022CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 1003 1022CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 1004 1023ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 1004 1023ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 1004 1023ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 1005 1024CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 1005 1024CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 1005 1024CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 1006 1025GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 1006 1025GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 1006 1025GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 1007 1026GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 1007 1026GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 1007 1026GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 1008 1027TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 1008 1027TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 1008 1027TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 1009 1028CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 1009 1028CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 1009 1028CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 1010 1029GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 1010 1029GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 1010 1029GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 1015 1034CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 1036 1055CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 1038 1057GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 1040 1059TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 1040 1059TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 1042 1061CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 1045 1064TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 1047 1066CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 1051 1070GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 1053 1072CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 1064 1083GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 410760 1088 1107CCGAATAAACTCCAGGCCTA 5-10-5 96.7 265 406045 1096 1115TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 1098 1117GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 1100 1119CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 1102 1121ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 1104 1123GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 1108 1127GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 1119 1138GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 1132 1151AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 1154 1173GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 1200 1219CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 1210 1229GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 1212 1231CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 1214 1233AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 1216 1235GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 1218 1237CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 1219 1238CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 279 405917 1222 1241TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 1224 1243AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 1226 1245GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 1228 1247CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 1230 1249CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 1232 1251GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 410765 1273 1292ATGACCTCGGGAGCTGAGGC 5-10-5 50.4 286 410766 1283 1302CCCAACTGTGATGACCTCGG 5-10-5 70.6 287 405923 1295 1314GGCATTGGTGGCCCCAACTG 5-10-5 94.3 288 410767 1305 1324GCTGGTCTTGGGCATTGGTG 5-10-5 65.1 289 405924 1318 1337CCCAGGGTCACCGGCTGGTC 5-10-5 93.1 290 410735 1320 1339TCCCCAGGGTCACCGGCTGG 5-10-5 78.1 291 405925 1322 1341AGTCCCCAGGGTCACCGGCT 5-10-5 93.5 292 405926 1324 1343AAAGTCCCCAGGGTCACCGG 5-10-5 94.6 293 405927 1328 1347CCCCAAAGTCCCCAGGGTCA 5-10-5 94.5 294 405928 1333 1352TTGGTCCCCAAAGTCCCCAG 5-10-5 90.0 295 405929 1337 1356AAAGTTGGTCCCCAAAGTCC 5-10-5 85.5 296 399895 1340 1359GCCAAAGTTGGTCCCCAAAG 3-14-3 84.6 36 395173 1340 1359GCCAAAGTTGGTCCCCAAAG 5-10-5 94.8 36 405930 1342 1361CGGCCAAAGTTGGTCCCCAA 5-10-5 92.9 297 405931 1344 1363AGCGGCCAAAGTTGGTCCCC 5-10-5 88.8 298 405932 1346 1365ACAGCGGCCAAAGTTGGTCC 5-10-5 90.6 299 405933 1348 1367ACACAGCGGCCAAAGTTGGT 5-10-5 NA 300 405934 1350 1369 CCACACAGCGGCCAAAGTTG5-10-5 92.7 301 405935 1354 1373 AGGTCCACACAGCGGCCAAA 5-10-5 86.0 302410736 1356 1375 AGAGGTCCACACAGCGGCCA 5-10-5 73.8 303 405936 1358 1377AAAGAGGTCCACACAGCGGC 5-10-5 93.9 304 410768 1380 1399CAATGATGTCCTCCCCTGGG 5-10-5 79.7 305 405937 1387 1406GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 1391 1410GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 1393 1412TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 1395 1414AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 1397 1416GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 1400 1419GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 1400 1419GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 1402 1421GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 1404 1423AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 1406 1425GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 1407 1426AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 1409 1428AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 1413 1432ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 1415 1434TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 1425 1444TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405948 1467 1486TCATGGCTGCAATGCCAGCC 5-10-5 45.9 319 399806 1470 1489GCATCATGGCTGCAATGCCA 5-10-5 82.8 42 399962 1470 1489GCATCATGGCTGCAATGCCA 3-14-3 86.1 42 405949 1472 1491CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 1474 1493GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 1476 1495CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 1480 1499TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 1482 1501GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 1484 1503CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 1486 1505TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 1500 1519CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 1513 1532CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 1515 1534GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 1519 1538ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 1521 1540GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 1523 1542GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 1525 1544AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 1528 1547GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 1530 1549CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 1532 1551 GGCAGAGAAGTGGATCAGTC5-10-5 59.7 337 405964 1536 1555 CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338405965 1541 1560 GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 1543 1562ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 1547 1566ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 1549 1568TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 1552 1571GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 1554 1573AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 1556 1575CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 1558 1577AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 1560 1579GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 1562 1581AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 1562 1581 AGGGAACCAGGCCTCATTGA2-13-5 NA 348 405887 1562 1581 AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 4105971563 1582 CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 1563 1582CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 1563 1582 CAGGGAACCAGGCCTCATTG2-13-5 NA 349 410674 1564 1583 TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49399812 1564 1583 TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 1564 1583TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 1565 1584CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 1565 1584CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 1565 1584CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 1566 1585CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 1566 1585CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 1566 1585CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 1567 1586TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 1567 1586TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 1567 1586TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 1568 1587GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 1568 1587GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 1568 1587GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 1569 1588GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 1569 1588GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 1569 1588GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 1570 1589TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 1570 1589TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 1570 1589TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 1571 1590CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 1571 1590CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 1571 1590CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 1572 1591GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 1572 1591GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 1572 1591GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 1573 1592CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 1578 1597GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 1580 1599CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 1583 1602GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 1583 1602GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 1628 1647ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 395181 1640 1659 AAACAGCTGCCAACCTGCCC5-10-5 87.5 52 405975 1642 1661 CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361405976 1647 1666 TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 1649 1668AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 410772 1660 1679GCTGACCATACAGTCCTGCA 5-10-5 91.2 364 405978 1672 1691GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 1675 1694GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 1677 1696GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 1679 1698CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 1681 1700ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 1683 1702CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 1685 1704GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 1687 1706GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 1735 1754CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 1737 1756AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 1742 1761GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 1744 1763CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 399905 1812 1831CGTTGTGGGCCCGGCAGACC 3-14-3 91.2 57 395183 1812 1831CGTTGTGGGCCCGGCAGACC 5-10-5 93.1 57 410541 1849 1868CACCTGGCAATGGCGTAGAC 5-10-5 46.1 376 410605 1849 1868CACCTGGCAATGGCGTAGAC 3-14-3 67.4 376 410683 1849 1868CACCTGGCAATGGCGTAGAC 2-13-5 71.8 376 410606 1850 1869GCACCTGGCAATGGCGTAGA 3-14-3 74.3 377 410542 1850 1869GCACCTGGCAATGGCGTAGA 5-10-5 75.5 377 410684 1850 1869GCACCTGGCAATGGCGTAGA 2-13-5 78.7 377 410543 1851 1870AGCACCTGGCAATGGCGTAG 5-10-5 76.4 378 410685 1851 1870AGCACCTGGCAATGGCGTAG 2-13-5 76.9 378 410607 1851 1870AGCACCTGGCAATGGCGTAG 3-14-3 77.1 378 410544 1852 1871CAGCACCTGGCAATGGCGTA 5-10-5 62.7 379 410608 1852 1871CAGCACCTGGCAATGGCGTA 3-14-3 69.6 379 410686 1852 1871CAGCACCTGGCAATGGCGTA 2-13-5 81.0 379 410545 1853 1872GCAGCACCTGGCAATGGCGT 5-10-5 75.5 380 410687 1853 1872GCAGCACCTGGCAATGGCGT 2-13-5 79.2 380 410609 1853 1872GCAGCACCTGGCAATGGCGT 3-14-3 83.2 380 410610 1854 1873GGCAGCACCTGGCAATGGCG 3-14-3 67.5 381 410688 1854 1873GGCAGCACCTGGCAATGGCG 2-13-5 89.3 381 405988 1854 1873GGCAGCACCTGGCAATGGCG 5-10-5 95.9 381 410546 1855 1874AGGCAGCACCTGGCAATGGC 5-10-5 74.8 382 410689 1855 1874AGGCAGCACCTGGCAATGGC 2-13-5 83.9 382 410611 1855 1874AGGCAGCACCTGGCAATGGC 3-14-3 88.2 382 410612 1856 1875CAGGCAGCACCTGGCAATGG 3-14-3 72.8 383 410690 1856 1875CAGGCAGCACCTGGCAATGG 2-13-5 74.8 383 405989 1856 1875CAGGCAGCACCTGGCAATGG 5-10-5 88.1 383 410547 1857 1876GCAGGCAGCACCTGGCAATG 5-10-5 63.0 384 410691 1857 1876GCAGGCAGCACCTGGCAATG 2-13-5 70.2 384 410613 1857 1876GCAGGCAGCACCTGGCAATG 3-14-3 76.7 384 395184 1858 1877AGCAGGCAGCACCTGGCAAT 5-10-5 68.5 58 410692 1858 1877AGCAGGCAGCACCTGGCAAT 2-13-5 69.1 58 399906 1858 1877AGCAGGCAGCACCTGGCAAT 3-14-3 94.2 58 410614 1859 1878TAGCAGGCAGCACCTGGCAA 3-14-3 36.6 385 410548 1859 1878TAGCAGGCAGCACCTGGCAA 5-10-5 75.0 385 410693 1859 1878TAGCAGGCAGCACCTGGCAA 2-13-5 82.0 385 405990 1860 1879GTAGCAGGCAGCACCTGGCA 5-10-5 96.8 386 410773 1905 1924TGGCCTCAGCTGGTGGAGCT 5-10-5 61.3 387 410549 1915 1934GTCCCCATGCTGGCCTCAGC 5-10-5 63.0 388 410615 1915 1934GTCCCCATGCTGGCCTCAGC 3-14-3 72.5 388 410694 1915 1934GTCCCCATGCTGGCCTCAGC 2-13-5 74.8 388 410550 1916 1935GGTCCCCATGCTGGCCTCAG 5-10-5 69.4 389 410616 1916 1935GGTCCCCATGCTGGCCTCAG 3-14-3 80.7 389 410695 1916 1935GGTCCCCATGCTGGCCTCAG 2-13-5 85.5 389 410551 1917 1936GGGTCCCCATGCTGGCCTCA 5-10-5 68.7 390 410696 1917 1936GGGTCCCCATGCTGGCCTCA 2-13-5 86.3 390 410617 1917 1936GGGTCCCCATGCTGGCCTCA 3-14-3 86.5 390 410552 1918 1937CGGGTCCCCATGCTGGCCTC 5-10-5 77.9 391 410618 1918 1937CGGGTCCCCATGCTGGCCTC 3-14-3 86.7 391 410697 1918 1937CGGGTCCCCATGCTGGCCTC 2-13-5 87.0 391 410553 1919 1938ACGGGTCCCCATGCTGGCCT 5-10-5 72.8 392 410619 1919 1938ACGGGTCCCCATGCTGGCCT 3-14-3 84.1 392 410698 1919 1938ACGGGTCCCCATGCTGGCCT 2-13-5 87.0 392 410699 1920 1939CACGGGTCCCCATGCTGGCC 2-13-5 79.5 59 395185 1920 1939CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 399907 1920 1939CACGGGTCCCCATGCTGGCC 3-14-3 93.7 59 410620 1921 1940ACACGGGTCCCCATGCTGGC 3-14-3 74.4 393 410554 1921 1940ACACGGGTCCCCATGCTGGC 5-10-5 79.7 393 410700 1921 1940ACACGGGTCCCCATGCTGGC 2-13-5 84.4 393 410621 1922 1941GACACGGGTCCCCATGCTGG 3-14-3 76.0 394 410701 1922 1941GACACGGGTCCCCATGCTGG 2-13-5 83.4 394 405991 1922 1941GACACGGGTCCCCATGCTGG 5-10-5 91.9 394 410555 1923 1942GGACACGGGTCCCCATGCTG 5-10-5 77.0 395 410622 1923 1942GGACACGGGTCCCCATGCTG 3-14-3 79.8 395 410702 1923 1942GGACACGGGTCCCCATGCTG 2-13-5 85.0 395 410703 1924 1943TGGACACGGGTCCCCATGCT 2-13-5 70.4 396 410623 1924 1943TGGACACGGGTCCCCATGCT 3-14-3 78.9 396 405992 1924 1943TGGACACGGGTCCCCATGCT 5-10-5 89.3 396 410704 1925 1944GTGGACACGGGTCCCCATGC 2-13-5 78.6 397 410556 1925 1944GTGGACACGGGTCCCCATGC 5-10-5 81.4 397 410624 1925 1944GTGGACACGGGTCCCCATGC 3-14-3 82.8 397 410625 1926 1945AGTGGACACGGGTCCCCATG 3-14-3 64.4 398 410705 1926 1945AGTGGACACGGGTCCCCATG 2-13-5 74.7 398 405993 1926 1945AGTGGACACGGGTCCCCATG 5-10-5 85.5 398 410706 1927 1946CAGTGGACACGGGTCCCCAT 2-13-5 70.8 399 410557 1927 1946CAGTGGACACGGGTCCCCAT 5-10-5 74.2 399 410626 1927 1946CAGTGGACACGGGTCCCCAT 3-14-3 76.8 399 410627 1928 1947GCAGTGGACACGGGTCCCCA 3-14-3 71.4 400 410707 1928 1947GCAGTGGACACGGGTCCCCA 2-13-5 72.8 400 405994 1928 1947GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 410708 1929 1948GGCAGTGGACACGGGTCCCC 2-13-5 79.0 401 410628 1929 1948GGCAGTGGACACGGGTCCCC 3-14-3 88.1 401 410558 1929 1948GGCAGTGGACACGGGTCCCC 5-10-5 88.9 401 410629 1930 1949TGGCAGTGGACACGGGTCCC 3-14-3 66.5 402 410709 1930 1949TGGCAGTGGACACGGGTCCC 2-13-5 66.9 402 405995 1930 1949TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 410630 1931 1950GTGGCAGTGGACACGGGTCC 3-14-3 54.1 403 410710 1931 1950GTGGCAGTGGACACGGGTCC 2-13-5 61.4 403 410559 1931 1950GTGGCAGTGGACACGGGTCC 5-10-5 77.5 403 410711 1932 1951GGTGGCAGTGGACACGGGTC 2-13-5 NA 404 410631 1932 1951 GGTGGCAGTGGACACGGGTC3-14-3 NA 404 410560 1932 1951 GGTGGCAGTGGACACGGGTC 5-10-5 42.5 404410712 1933 1952 TGGTGGCAGTGGACACGGGT 2-13-5 NA 405 410632 1933 1952TGGTGGCAGTGGACACGGGT 3-14-3 NA 405 410561 1933 1952 TGGTGGCAGTGGACACGGGT5-10-5 NA 405 410774 1936 1955 TGTTGGTGGCAGTGGACACG 5-10-5 47.8 406410775 1962 1981 AGCTGCAGCCTGTGAGGACG 5-10-5 36.3 407 410776 1990 2009GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 2010 2029TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 2040 2059CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 2100 2119CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 2100 2119CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 2100 2119CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 2101 2120ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 2101 2120ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 2101 2120ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 2102 2121GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 2102 2121GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 2102 2121GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 2103 2122TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 2103 2122TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 2103 2122TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 2104 2123TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 2104 2123TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 2104 2123TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 2105 2124CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 2105 2124CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 2105 2124CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 2107 2126TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 2120 2139CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410780 2150 2169GCAGGCCACGGTCACCTGCT 5-10-5 60.9 417 410781 2187 2206GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 2305 2324CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 2305 2324CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 2305 2324CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 2306 2325GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 2306 2325GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 2306 2325GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 2307 2326AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 2307 2326AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 2307 2326AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 2308 2327CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 2308 2327CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 2308 2327CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 2309 2328GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 2309 2328GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 2309 2328GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 2310 2329GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 2310 2329GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 2310 2329GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 2311 2330CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 2311 2330CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410570 2311 2330CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 2312 2331CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 2312 2331CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 2312 2331CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 2313 2332TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 2313 2332TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 2313 2332TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 410572 2314 2333CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 2314 2333CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 2314 2333CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 2315 2334GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 2315 2334GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 2315 2334GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 2325 2344CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 2335 2354GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 2509 2528CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 2597 2616ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 2828 2847ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 2829 2848GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 2830 2849AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 2831 2850GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 2832 2851GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 2832 2851GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 2833 2852TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 2834 2853CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 2835 2854GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 2836 2855AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 2902 2921GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 2903 2922GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 2904 2923TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 2905 2924CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 2906 2925TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 2907 2926CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 2908 2927CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 2909 2928GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 2910 2929AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 2988 3007AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition ofPCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited atleast 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155,395158, 395163, 395165, 395166, 395168, 395173, 395181, 395183, 395185,395186, 395187, 395194, 399798, 399801, 399804, 399806, 399817, 399819,399821, 399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904,399905, 399906, 399907, 399912, 399916, 399954, 399960, 399962, 399967,399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865,405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876,405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889,405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899,405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908,405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921,405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930,405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944,405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960,405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982,405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994,405995, 405996, 405997, 406002, 406003, 406005, 406008, 406009, 406014,406016, 406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032,406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043,406044, 406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589,410590, 410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617,410618, 410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650,410652, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686,410688, 410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701,410702, 410713, 410714, 410715, 410716, 410723, 410724, 410730, 410751,410754, 410757, 410759, 410760, 410769, and 410772. The target segmentsto which these antisense oligonucleotides are targeted are active targetsegments. The target regions to which these antisense oligonucleotidesare targeted are active target regions.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395181,395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821,399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906,399907, 399916, 399954, 399962, 399969, 399978, 405861, 405862, 405864,405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879,405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894,405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904,405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916,405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928,405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952,405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977,405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991,405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014,406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035,406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558,410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628,410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698,410702, 410716, 410724, 410730, 410757, 410760, and 410772 eachexhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183,395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399877,399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907,399916, 399954, 399969, 399978, 405861, 405862, 405864, 405873, 405874,405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884,405891, 405893, 405894, 405895, 405899, 405903, 405904, 405905, 405906,405907, 405910, 405913, 405914, 405916, 405923, 405924, 405925, 405926,405927, 405928, 405930, 405932, 405934, 405936, 405941, 405953, 405955,405956, 405959, 405960, 405978, 405988, 405990, 405991, 405994, 405995,405996, 405997, 406008, 406009, 406014, 406023, 406028, 406029, 406030,406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592,410647, 410668, 410760, and 410772 each exhibited at least 90%inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978,405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, 406044,and 410760 each exhibited at least 95% inhibition of PCSK9 mRNA levels.

TABLE 18 Antisense inhibition of PCSK9 in human cells (Hep3B) targetingSEQ ID NO: 2 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % IDIsis No. NO: 2 NO: 2 Sequence 5′-3′ Motif inhibition NO: 410742 24332452 GCCTGGAGCTGACGGTGCCC 5-10-5 75.1 159 405999 2437 2456GACCGCCTGGAGCTGACGGT 5-10-5 70.3 160 395151 2439 2458AGGACCGCCTGGAGCTGACG 5-10-5 57.3 6 405861 2545 2564 AAGGCTAGCACCAGCTCCTC5-10-5 90.5 162 405862 2546 2565 CAAGGCTAGCACCAGCTCCT 5-10-5 92.3 163405863 2547 2566 GCAAGGCTAGCACCAGCTCC 5-10-5 82.6 164 405864 2548 2567CGCAAGGCTAGCACCAGCTC 5-10-5 91.8 165 395152 2549 2568ACGCAAGGCTAGCACCAGCT 5-10-5 93.3 7 405865 2550 2569 AACGCAAGGCTAGCACCAGC5-10-5 84.6 166 405866 2551 2570 GAACGCAAGGCTAGCACCAG 5-10-5 81.8 167405867 2552 2571 GGAACGCAAGGCTAGCACCA 5-10-5 74.0 168 405868 2553 2572CGGAACGCAAGGCTAGCACC 5-10-5 76.8 169 399793 2556 2575CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 410743 2560 2579 TCCTCCTCGGAACGCAAGGC5-10-5 76.7 170 410744 2585 2604 GTGCTCGGGTGCTTCGGCCA 5-10-5 76.9 171410745 2605 2624 TGGAAGGTGGCTGTGGTTCC 5-10-5 41.7 172 395153 2619 2638CCTTGGCGCAGCGGTGGAAG 5-10-5 81.1 9 410746 6444 6463 CGTAGGTGCCAGGCAACCTC5-10-5 44.8 176 410747 6482 6501 TGACTGCGAGAGGTGGGTCT 5-10-5 NA 177406003 6492 6511 CAGTGCGCTCTGACTGCGAG 5-10-5 84.2 178 406004 6494 6513GGCAGTGCGCTCTGACTGCG 5-10-5 56.4 179 406005 6496 6515CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 406006 6499 6518CGGCGGGCAGTGCGCTCTGA 5-10-5 71.0 181 410748 6528 6547ATCCCCGGCGGGCAGCCTGG 5-10-5 56.0 182 406007 6532 6551AGGTATCCCCGGCGGGCAGC 5-10-5 76.3 183 410574 6534 6553TGAGGTATCCCCGGCGGGCA 3-14-3 69.1 184 410529 6534 6553TGAGGTATCCCCGGCGGGCA 5-10-5 77.4 184 410647 6534 6553TGAGGTATCCCCGGCGGGCA 2-13-5 90.7 184 410575 6535 6554GTGAGGTATCCCCGGCGGGC 3-14-3 71.5 185 410648 6535 6554GTGAGGTATCCCCGGCGGGC 2-13-5 82.8 185 410530 6535 6554GTGAGGTATCCCCGGCGGGC 5-10-5 83.2 185 410649 6536 6555GGTGAGGTATCCCCGGCGGG 2-13-5 53.5 186 410576 6536 6555GGTGAGGTATCCCCGGCGGG 3-14-3 56.2 186 410531 6536 6555GGTGAGGTATCCCCGGCGGG 5-10-5 63.6 186 410650 6537 6556TGGTGAGGTATCCCCGGCGG 2-13-5 86.1 11 399877 6537 6556TGGTGAGGTATCCCCGGCGG 3-14-3 92.6 11 395155 6537 6556TGGTGAGGTATCCCCGGCGG 5-10-5 92.7 11 410577 6538 6557TTGGTGAGGTATCCCCGGCG 3-14-3 62.4 187 410532 6538 6557TTGGTGAGGTATCCCCGGCG 5-10-5 63.5 187 410651 6538 6557TTGGTGAGGTATCCCCGGCG 2-13-5 75.5 187 410652 6539 6558CTTGGTGAGGTATCCCCGGC 2-13-5 81.2 188 410578 6539 6558CTTGGTGAGGTATCCCCGGC 3-14-3 83.3 188 406008 6539 6558CTTGGTGAGGTATCCCCGGC 5-10-5 95.2 188 410653 6540 6559TCTTGGTGAGGTATCCCCGG 2-13-5 68.9 189 410579 6540 6559TCTTGGTGAGGTATCCCCGG 3-14-3 74.0 189 410533 6540 6559TCTTGGTGAGGTATCCCCGG 5-10-5 79.0 189 410580 6541 6560ATCTTGGTGAGGTATCCCCG 3-14-3 62.3 190 410654 6541 6560ATCTTGGTGAGGTATCCCCG 2-13-5 65.2 190 406009 6541 6560ATCTTGGTGAGGTATCCCCG 5-10-5 90.7 190 410534 6542 6561GATCTTGGTGAGGTATCCCC 5-10-5 59.8 191 410581 6542 6561GATCTTGGTGAGGTATCCCC 3-14-3 72.9 191 410655 6542 6561GATCTTGGTGAGGTATCCCC 2-13-5 77.4 191 399794 6543 6562GGATCTTGGTGAGGTATCCC 5-10-5 56.5 12 410656 6543 6562GGATCTTGGTGAGGTATCCC 2-13-5 68.0 12 399950 6543 6562GGATCTTGGTGAGGTATCCC 3-14-3 74.4 12 410535 6544 6563AGGATCTTGGTGAGGTATCC 5-10-5 65.7 192 410582 6544 6563AGGATCTTGGTGAGGTATCC 3-14-3 69.5 192 410657 6544 6563AGGATCTTGGTGAGGTATCC 2-13-5 73.1 192 406010 6546 6565GCAGGATCTTGGTGAGGTAT 5-10-5 56.9 193 406011 6548 6567ATGCAGGATCTTGGTGAGGT 5-10-5 70.7 194 406012 6550 6569ACATGCAGGATCTTGGTGAG 5-10-5 69.1 195 406013 6554 6573GAAGACATGCAGGATCTTGG 5-10-5 77.4 196 395156 6557 6576ATGGAAGACATGCAGGATCT 5-10-5 76.7 14 410749 6565 6584AGAAGGCCATGGAAGACATG 5-10-5 59.4 197 410750 6575 6594GAAGCCAGGAAGAAGGCCAT 5-10-5 57.5 198 399879 6583 6602TTCACCAGGAAGCCAGGAAG 3-14-3 91.9 15 406014 6585 6604TCTTCACCAGGAAGCCAGGA 5-10-5 94.0 199 406015 6590 6609ACTCATCTTCACCAGGAAGC 5-10-5 78.8 200 406016 6592 6611CCACTCATCTTCACCAGGAA 5-10-5 87.2 201 410730 6594 6613CGCCACTCATCTTCACCAGG 5-10-5 85.4 202 406017 6596 6615GTCGCCACTCATCTTCACCA 5-10-5 76.3 203 406018 6598 6617AGGTCGCCACTCATCTTCAC 5-10-5 63.2 204 406019 6600 6619GCAGGTCGCCACTCATCTTC 5-10-5 68.1 205 406020 6602 6621CAGCAGGTCGCCACTCATCT 5-10-5 83.7 206 410731 6604 6623TCCAGCAGGTCGCCACTCAT 5-10-5 72.9 207 395158 9130 9149CCTCGATGTAGTCGACATGG 5-10-5 93.9 17 410752 9149 9168GCAAAGACAGAGGAGTCCTC 5-10-5 76.3 209 406021 9207 9226TTCATCCGCCCGGTACCGTG 5-10-5 79.6 210 406022 9209 9228TATTCATCCGCCCGGTACCG 5-10-5 78.7 211 406023 9212 9231TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 406024 9214 9233GCTGGTATTCATCCGCCCGG 5-10-5 69.7 213 410732 9216 9235GGGCTGGTATTCATCCGCCC 5-10-5 30.4 214 399935 14601 14620AGGACCCAAGTCATCCTGCT 3-14-3 81.1 96 399936 14631 14650GGCCATCAGCTGGCAATGCT 3-14-3 89.0 124 410753 14877 14896ATACACCTCCACCAGGCTGC 5-10-5 72.8 315 406025 14888 14907GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 14893 14912TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 14918 14937TCGATTTCCCGGTGGTCACT 5-10-5 87.6 218 405870 14919 14938CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219 405871 14920 14939CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 14921 14940CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 14922 14941GCCCTCGATTTCCCGGTGGT 5-10-5 84.3 22 399954 14922 14941GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873 14923 14942TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 14924 14943CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 14925 14944CCTGCCCTCGATTTCCCGGT 5-10-5 93.5 224 405876 14926 14945CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225 406027 14930 14949ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 14932 14951CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 14934 14953GACCATGACCCTGCCCTCGA 5-10-5 91.4 228 406030 14938 14957CGGTGACCATGACCCTGCCC 5-10-5 95.6 229 406031 14940 14959GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 14942 14961AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 14944 14963CGAAGTCGGTGACCATGACC 5-10-5 88.5 232 410754 14954 14973GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233 395163 14979 14998GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410756 15254 15273GGTGGGTGCCATGACTGTCA 5-10-5 67.1 235 406033 15261 15280CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 406034 15266 15285CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 15271 15290GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 15279 15298TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 15283 15302GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 15286 15305GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 15291 15310GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 15291 15310GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 15291 15310GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 15292 15311GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 15292 15311GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 15292 15311GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 15293 15312TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 15293 15312TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 15293 15312TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 15294 15313TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 15294 15313TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 15294 15313TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 15295 15314CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 15295 15314CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 15295 15314CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 15296 15315CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 15296 15315CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 15296 15315CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 15297 15316CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 15297 15316CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 15297 15316CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 15298 15317ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 15298 15317ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 15298 15317ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 15299 15318CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 15299 15318CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 15299 15318CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 15300 15319GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 15300 15319GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 15300 15319GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 15301 15320GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 15301 15320GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 15301 15320GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 15302 15321TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 15302 15321TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 15302 15321TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 15303 15322CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 15303 15322CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 15303 15322CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 15304 15323GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 15304 15323GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 15304 15323GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 15309 15328CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 15330 15349CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 15332 15351GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 15334 15353TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 15334 15353TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 15336 15355CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 15339 15358TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 15341 15360CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 15345 15364GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 15347 15366CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 15358 15377GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 406045 18591 18610TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 18593 18612GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 18595 18614CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 18597 18616ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 18599 18618GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 18603 18622GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 18614 18633GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 18627 18646AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 18649 18668GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 18695 18714CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 18705 18724GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 18707 18726CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 18709 18728AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 18711 18730GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 18713 18732CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 18714 18733CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 279 405917 18717 18736TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 18719 18738AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 18721 18740GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 18723 18742CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 18725 18744CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 18727 18746GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 405923 19931 19950GGCATTGGTGGCCCCAACTG 5-10-5 94.3 288 410767 19941 19960GCTGGTCTTGGGCATTGGTG 5-10-5 65.1 289 405924 19954 19973CCCAGGGTCACCGGCTGGTC 5-10-5 93.1 290 410735 19956 19975TCCCCAGGGTCACCGGCTGG 5-10-5 78.1 291 405925 19958 19977AGTCCCCAGGGTCACCGGCT 5-10-5 93.5 292 405926 19960 19979AAAGTCCCCAGGGTCACCGG 5-10-5 94.6 293 405927 19964 19983CCCCAAAGTCCCCAGGGTCA 5-10-5 94.5 294 405928 19969 19988TTGGTCCCCAAAGTCCCCAG 5-10-5 90.0 295 405929 19973 19992AAAGTTGGTCCCCAAAGTCC 5-10-5 85.5 296 399895 19976 19995GCCAAAGTTGGTCCCCAAAG 3-14-3 84.6 36 395173 19976 19995GCCAAAGTTGGTCCCCAAAG 5-10-5 94.8 36 405930 19978 19997CGGCCAAAGTTGGTCCCCAA 5-10-5 92.9 297 405931 19980 19999AGCGGCCAAAGTTGGTCCCC 5-10-5 88.8 298 405932 19982 20001ACAGCGGCCAAAGTTGGTCC 5-10-5 90.6 299 405933 19984 20003ACACAGCGGCCAAAGTTGGT 5-10-5 NA 300 405934 19986 20005CCACACAGCGGCCAAAGTTG 5-10-5 92.7 301 405935 19990 20009AGGTCCACACAGCGGCCAAA 5-10-5 86.0 302 410736 19992 20011AGAGGTCCACACAGCGGCCA 5-10-5 73.8 303 405936 19994 20013AAAGAGGTCCACACAGCGGC 5-10-5 93.9 304 410768 20016 20035CAATGATGTCCTCCCCTGGG 5-10-5 79.7 305 405937 20023 20042GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 20027 20046GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 20029 20048TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 20031 20050AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 20033 20052GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 20036 20055GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 20036 20055GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 20038 20057GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 20040 20059AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 20042 20061GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 20043 20062AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 20045 20064AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 20049 20068ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 20051 20070TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 20061 20080TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405949 20629 20648CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 20631 20650GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 20633 20652CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 20637 20656TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 20639 20658GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 20641 20660CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 20643 20662TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 20657 20676CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 20670 20689CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 20672 20691GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 20676 20695ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 20678 20697GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 20680 20699GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 20682 20701AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 20685 20704GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 20687 20706CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 20689 20708GGCAGAGAAGTGGATCAGTC 5-10-5 59.7 337 405964 20693 20712CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338 405965 20698 20717GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 20700 20719ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 20704 20723ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 20706 20725TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 20709 20728GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 20711 20730AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 20713 20732CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 20715 20734AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 20717 20736GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 20719 20738AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 20719 20738AGGGAACCAGGCCTCATTGA 2-13-5 NA 348 405887 20719 20738AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 410597 20720 20739CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 20720 20739CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 20720 20739CAGGGAACCAGGCCTCATTG 2-13-5 NA 349 410674 20721 20740TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49 399812 20721 20740TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 20721 20740TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 20722 20741CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 20722 20741CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 20722 20741CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 20723 20742CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 20723 20742CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 20723 20742CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 20724 20743TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 20724 20743TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 20724 20743TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 20725 20744GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 20725 20744GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 20725 20744GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 20726 20745GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 20726 20745GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 20726 20745GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 20727 20746TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 20727 20746TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 20727 20746TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 20728 20747CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 20728 20747CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 20728 20747CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 20729 20748GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 20729 20748GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 20729 20748GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 20730 20749CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 20735 20754GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 20737 20756CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 20740 20759GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 20740 20759GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 20785 20804ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 405975 21088 21107CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361 405976 21093 21112TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 21095 21114AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 410772 21106 21125GCTGACCATACAGTCCTGCA 5-10-5 91.2 264 405978 21118 21137GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 21121 21140GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 21123 21142GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 21125 21144CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 21127 21146ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 21129 21148CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 21131 21150GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 21133 21152GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 21181 21200CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 21183 21202AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 21188 21207GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 21190 21209CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 399868 21696 21715GGCACTGCCCTTCCACCAAA 5-10-5 85.8 123 399905 22096 22115CGTTGTGGGCCCGGCAGACC 3-14-3 91.2 57 395183 22096 22115CGTTGTGGGCCCGGCAGACC 5-10-5 93.1 57 410541 22133 22152CACCTGGCAATGGCGTAGAC 5-10-5 46.1 376 410605 22133 22152CACCTGGCAATGGCGTAGAC 3-14-3 67.4 376 410683 22133 22152CACCTGGCAATGGCGTAGAC 2-13-5 71.8 376 410606 22134 22153GCACCTGGCAATGGCGTAGA 3-14-3 74.3 377 410542 22134 22153GCACCTGGCAATGGCGTAGA 5-10-5 75.5 377 410684 22134 22153GCACCTGGCAATGGCGTAGA 2-13-5 78.7 377 410543 22135 22154AGCACCTGGCAATGGCGTAG 5-10-5 76.4 378 410685 22135 22154AGCACCTGGCAATGGCGTAG 2-13-5 76.9 378 410607 22135 22154AGCACCTGGCAATGGCGTAG 3-14-3 77.1 378 410544 22136 22155CAGCACCTGGCAATGGCGTA 5-10-5 62.7 379 410608 22136 22155CAGCACCTGGCAATGGCGTA 3-14-3 69.6 379 410686 22136 22155CAGCACCTGGCAATGGCGTA 2-13-5 81.0 379 410545 22137 22156GCAGCACCTGGCAATGGCGT 5-10-5 75.5 380 410687 22137 22156GCAGCACCTGGCAATGGCGT 2-13-5 79.2 380 410609 22137 22156GCAGCACCTGGCAATGGCGT 3-14-3 83.2 380 410610 22138 22157GGCAGCACCTGGCAATGGCG 3-14-3 67.5 381 410688 22138 22157GGCAGCACCTGGCAATGGCG 2-13-5 89.3 381 405988 22138 22157GGCAGCACCTGGCAATGGCG 5-10-5 95.9 381 410546 22139 22158AGGCAGCACCTGGCAATGGC 5-10-5 74.8 382 410689 22139 22158AGGCAGCACCTGGCAATGGC 2-13-5 83.9 382 410611 22139 22158AGGCAGCACCTGGCAATGGC 3-14-3 88.2 382 410612 22140 22159CAGGCAGCACCTGGCAATGG 3-14-3 72.8 383 410690 22140 22159CAGGCAGCACCTGGCAATGG 2-13-5 74.8 383 405989 22140 22159CAGGCAGCACCTGGCAATGG 5-10-5 88.1 383 410547 22141 22160GCAGGCAGCACCTGGCAATG 5-10-5 63.0 384 410691 22141 22160GCAGGCAGCACCTGGCAATG 2-13-5 70.2 384 410613 22141 22160GCAGGCAGCACCTGGCAATG 3-14-3 76.7 384 395184 22142 22161AGCAGGCAGCACCTGGCAAT 5-10-5 68.5 58 410692 22142 22161AGCAGGCAGCACCTGGCAAT 2-13-5 69.1 58 399906 22142 22161AGCAGGCAGCACCTGGCAAT 3-14-3 94.2 58 410614 22143 22162TAGCAGGCAGCACCTGGCAA 3-14-3 36.6 385 410548 22143 22162TAGCAGGCAGCACCTGGCAA 5-10-5 75.0 385 410693 22143 22162TAGCAGGCAGCACCTGGCAA 2-13-5 82.0 385 405990 22144 22163GTAGCAGGCAGCACCTGGCA 5-10-5 96.8 386 410773 22189 22208TGGCCTCAGCTGGTGGAGCT 5-10-5 61.3 387 410549 22199 22218GTCCCCATGCTGGCCTCAGC 5-10-5 63.0 388 410615 22199 22218GTCCCCATGCTGGCCTCAGC 3-14-3 72.5 388 410694 22199 22218GTCCCCATGCTGGCCTCAGC 2-13-5 74.8 388 410550 22200 22219GGTCCCCATGCTGGCCTCAG 5-10-5 69.4 389 410616 22200 22219GGTCCCCATGCTGGCCTCAG 3-14-3 80.7 389 410695 22200 22219GGTCCCCATGCTGGCCTCAG 2-13-5 85.5 389 410551 22201 22220GGGTCCCCATGCTGGCCTCA 5-10-5 68.7 390 410696 22201 22220GGGTCCCCATGCTGGCCTCA 2-13-5 86.3 390 410617 22201 22220GGGTCCCCATGCTGGCCTCA 3-14-3 86.5 390 410552 22202 22221CGGGTCCCCATGCTGGCCTC 5-10-5 77.9 391 410618 22202 22221CGGGTCCCCATGCTGGCCTC 3-14-3 86.7 391 410697 22202 22221CGGGTCCCCATGCTGGCCTC 2-13-5 87.0 391 410553 22203 22222ACGGGTCCCCATGCTGGCCT 5-10-5 72.8 392 410619 22203 22222ACGGGTCCCCATGCTGGCCT 3-14-3 84.1 392 410698 22203 22222ACGGGTCCCCATGCTGGCCT 2-13-5 87.0 392 410699 22204 22223CACGGGTCCCCATGCTGGCC 2-13-5 79.5 59 395185 22204 22223CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 399907 22204 22223CACGGGTCCCCATGCTGGCC 3-14-3 93.7 59 410620 22205 22224ACACGGGTCCCCATGCTGGC 3-14-3 74.4 393 410554 22205 22224ACACGGGTCCCCATGCTGGC 5-10-5 79.7 393 410700 22205 22224ACACGGGTCCCCATGCTGGC 2-13-5 84.4 393 410621 22206 22225GACACGGGTCCCCATGCTGG 3-14-3 76.0 394 410701 22206 22225GACACGGGTCCCCATGCTGG 2-13-5 83.4 394 405991 22206 22225GACACGGGTCCCCATGCTGG 5-10-5 91.9 394 410555 22207 22226GGACACGGGTCCCCATGCTG 5-10-5 77.0 395 410622 22207 22226GGACACGGGTCCCCATGCTG 3-14-3 79.8 395 410702 22207 22226GGACACGGGTCCCCATGCTG 2-13-5 85.0 395 410703 22208 22227TGGACACGGGTCCCCATGCT 2-13-5 70.4 396 410623 22208 22227TGGACACGGGTCCCCATGCT 3-14-3 78.9 396 405992 22208 22227TGGACACGGGTCCCCATGCT 5-10-5 89.3 396 410704 22209 22228GTGGACACGGGTCCCCATGC 2-13-5 78.6 397 410556 22209 22228GTGGACACGGGTCCCCATGC 5-10-5 81.4 397 410624 22209 22228GTGGACACGGGTCCCCATGC 3-14-3 82.8 397 410625 22210 22229AGTGGACACGGGTCCCCATG 3-14-3 64.4 398 410705 22210 22229AGTGGACACGGGTCCCCATG 2-13-5 74.7 398 405993 22210 22229AGTGGACACGGGTCCCCATG 5-10-5 85.5 398 410706 22211 22230CAGTGGACACGGGTCCCCAT 2-13-5 70.8 399 410557 22211 22230CAGTGGACACGGGTCCCCAT 5-10-5 74.2 399 410626 22211 22230CAGTGGACACGGGTCCCCAT 3-14-3 76.8 399 410627 22212 22231GCAGTGGACACGGGTCCCCA 3-14-3 71.4 400 410707 22212 22231GCAGTGGACACGGGTCCCCA 2-13-5 72.8 400 405994 22212 22231GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 410708 22213 22232GGCAGTGGACACGGGTCCCC 2-13-5 79.0 401 410628 22213 22232GGCAGTGGACACGGGTCCCC 3-14-3 88.1 401 410558 22213 22232GGCAGTGGACACGGGTCCCC 5-10-5 88.9 401 410629 22214 22233TGGCAGTGGACACGGGTCCC 3-14-3 66.5 402 410709 22214 22233TGGCAGTGGACACGGGTCCC 2-13-5 66.9 402 405995 22214 22233TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 410630 22215 22234GTGGCAGTGGACACGGGTCC 3-14-3 54.1 403 410710 22215 22234GTGGCAGTGGACACGGGTCC 2-13-5 61.4 403 410559 22215 22234GTGGCAGTGGACACGGGTCC 5-10-5 77.5 403 410711 22216 22235GGTGGCAGTGGACACGGGTC 2-13-5 NA 404 410631 22216 22235GGTGGCAGTGGACACGGGTC 3-14-3 NA 404 410560 22216 22235GGTGGCAGTGGACACGGGTC 5-10-5 42.5 404 410712 22217 22236TGGTGGCAGTGGACACGGGT 2-13-5 NA 405 410632 22217 22236TGGTGGCAGTGGACACGGGT 3-14-3 NA 405 410561 22217 22236TGGTGGCAGTGGACACGGGT 5-10-5 NA 405 410774 22220 22239TGTTGGTGGCAGTGGACACG 5-10-5 47.8 406 410776 23985 24004GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 24005 24024TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 24035 24054CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 24095 24114CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 24095 24114CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 24095 24114CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 24096 24115ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 24096 24115ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 24096 24115ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 24097 24116GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 24097 24116GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 24097 24116GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 24098 24117TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 24098 24117TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 24098 24117TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 24099 24118TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 24099 24118TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 24099 24118TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 24100 24119CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 24100 24119CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 24100 24119CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 24102 24121TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 24115 24134CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410781 25994 26013GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 26112 26131CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 26112 26131CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 26112 26131CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 26113 26132GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 26113 26132GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 26113 26132GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 26114 26133AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 26114 26133AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 26114 26133AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 26115 26134CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 26115 26134CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 26115 26134CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 26116 26135GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 26116 26135GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 26116 26135GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 26117 26136GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 26117 26136GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 26117 26136GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 26118 26137CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 26118 26137CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410570 26118 26137CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 26119 26138CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 26119 26138CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 26119 26138CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 26120 26139TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 26120 26139TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 26120 26139TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 410572 26121 26140CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 26121 26140CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 26121 26140CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 26122 26141GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 26122 26141GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 26122 26141GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 26132 26151CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 26142 26161GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 26316 26335CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 26404 26423ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 26635 26654ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 26636 26655GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 26637 26656AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 26638 26657GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 26639 26658GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 26639 26658GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 26640 26659TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 26641 26660CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 26642 26661GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 26643 26662AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 26709 26728GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 26710 26729GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 26711 26730TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 26712 26731CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 26713 26732TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 26714 26733CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 26715 26734CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 26716 26735GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 26717 26736AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 26795 26814AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition ofPCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited atleast 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155,395158, 395163, 395165, 395166, 395168, 395173, 395183, 395185, 395186,395187, 395194, 399798, 399801, 399804, 399817, 399819, 399821, 399868,399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904, 399905,399906, 399907, 399912, 399916, 399935, 399936, 399954, 399960, 399967,399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865,405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876,405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889,405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899,405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908,405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921,405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930,405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944,405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960,405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982,405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994,405995, 405996, 405997, 406003, 406005, 406008, 406009, 406014, 406016,406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032, 406033,406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044,406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589, 410590,410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617, 410618,410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650, 410652,410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686, 410688,410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701, 410702,410713, 410714, 410715, 410716, 410723, 410724, 410730, 410754, 410757,410759, 410769, and 410772. The target segments to which these antisenseoligonucleotides are targeted are active target segments. The targetregions to which these antisense oligonucleotides are targeted areactive target regions.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183,395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399868,399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906,399907, 399916, 399936, 399954, 399969, 399978, 405861, 405862, 405864,405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879,405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894,405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904,405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916,405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928,405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952,405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977,405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991,405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014,406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035,406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558,410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628,410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698,410702, 410716, 410724, 410730, 410757, and 410772 each exhibited atleast 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183,395185, 395186, 395187, 399801, 399817, 399819, 399821, 399877, 399879,399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916,399969, 399978, 405862, 405864, 405874, 405875, 405878, 405879, 405880,405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899,405903, 405906, 405910, 405913, 405916, 405923, 405924, 405925, 405926,405927, 405930, 405934, 405936, 405953, 405955, 405956, 405959, 405978,405988, 405990, 405991, 405994, 405995, 405996, 405997, 406008, 406014,406023, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041,406042, 406043, 406044, 410592, 410647, 410668, and 410772 eachexhibited at least 90% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978,405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, and406044 each exhibited at least 95% inhibition of PCSK9 mRNA levels.

TABLE 19 Antisense inhibition of PCSK9 in human cells (Hep3B) targetingSEQ ID NO: 3 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % IDIsis No. NO: 3 NO: 3 Sequence 5′-3′ Motif inhibition NO: 399935 10751094 AGGACCCAAGTCATCCTGCT 3-14-3 81.1 96 399936 1105 1124GGCCATCAGCTGGCAATGCT 3-14-3 89.0 124 410753 1351 1370ATACACCTCCACCAGGCTGC 5-10-5 72.8 215 406025 1362 1381GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 1367 1386TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 1392 1411TCGATTTCCCGGTGGTCACT 5-10-5 87.6 218 405870 1393 1412CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219 405871 1394 1413CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 1395 1414CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 1396 1415GCCCTCGATTTCCCGGTGGT 5-10-5 84.3 22 399954 1396 1415GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873 1397 1416TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 1398 1417CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 1399 1418CCTGCCCTCGATTTCCCGGT 5-10-5 93.5 224 405876 1400 1419CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225 406027 1404 1423ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 1406 1425CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 1408 1427GACCATGACCCTGCCCTCGA 5-10-5 91.4 228 406030 1412 1431CGGTGACCATGACCCTGCCC 5-10-5 95.6 229 406031 1414 1433GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 1416 1435AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 1418 1437CGAAGTCGGTGACCATGACC 5-10-5 88.5 232 410754 1428 1447GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233 395163 1453 1472GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410755 1463 1482TGGCCTGTCTGTGGAAGCGG 5-10-5 NA 234 410756 1490 1509 GGTGGGTGCCATGACTGTCA5-10-5 67.1 235 406033 1497 1516 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237406034 1502 1521 CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 1507 1526GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 1515 1534TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 1519 1538GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 1522 1541GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 1527 1546GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 1527 1546GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 1527 1546GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 1528 1547GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 1528 1547GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 1528 1547GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 1529 1548TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 1529 1548TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 1529 1548TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 1530 1549TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 1530 1549TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 1530 1549TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 1531 1550CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 1531 1550CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 1531 1550CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 1532 1551CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 1532 1551CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 1532 1551CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 1533 1552CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 1533 1552CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 1533 1552CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 1534 1553ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 1534 1553ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 1534 1553ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 1535 1554CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 1535 1554CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 1535 1554CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 1536 1555GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 1536 1555GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 1536 1555GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 1537 1556GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 1537 1556GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 1537 1556GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 1538 1557TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 1538 1557TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 1538 1557TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 1539 1558CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 1539 1558CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 1539 1558CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 1540 1559GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 1540 1559GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 1540 1559GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 1545 1564CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 1566 1585CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 1568 1587GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 1570 1589TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 1570 1589TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 1572 1591CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 1575 1594TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 1577 1596CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 1581 1600GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 1583 1602CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 1594 1613GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 410760 1618 1637CCGAATAAACTCCAGGCCTA 5-10-5 96.7 265 406045 1626 1645TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 1628 1647GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 1630 1649CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 1632 1651ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 1634 1653GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 1638 1657GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 1649 1668GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 1662 1681AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 1684 1703GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 1730 1749CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 1740 1759GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 1742 1761CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 1744 1763AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 1746 1765GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 1748 1767CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 1749 1768CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 249 405917 1752 1771TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 1754 1773AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 1756 1775GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 1758 1777CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 1760 1779CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 1762 1781GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 405937 1820 1839GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 1824 1843GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 1826 1845TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 1828 1847AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 1830 1849GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 1833 1852GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 1833 1852GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 1835 1854GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 1837 1856AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 1839 1858GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 1840 1859AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 1842 1861AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 1846 1865ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 1848 1867TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 1858 1877TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405948 1900 1919TCATGGCTGCAATGCCAGCC 5-10-5 45.9 319 399806 1903 1922GCATCATGGCTGCAATGCCA 5-10-5 82.8 42 399962 1903 1922GCATCATGGCTGCAATGCCA 3-14-3 86.1 42 405949 1905 1924CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 1907 1926GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 1909 1928CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 1913 1932TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 1915 1934GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 1917 1936CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 1919 1938TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 1933 1952CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 1946 1965CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 1948 1967GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 1952 1971ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 1954 1973GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 1956 1975GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 1958 1977AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 1961 1980GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 1963 1982CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 1965 1984 GGCAGAGAAGTGGATCAGTC5-10-5 59.7 337 405964 1969 1988 CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338405965 1974 1993 GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 1976 1995ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 1980 1999ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 1982 2001TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 1985 2004GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 1987 2006AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 1989 2008CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 1991 2010AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 1993 2012GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 1995 2014AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 1995 2014 AGGGAACCAGGCCTCATTGA2-13-5 NA 348 405887 1995 2014 AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 4105971996 2015 CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 1996 2015CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 1996 2015 CAGGGAACCAGGCCTCATTG2-13-5 NA 349 410674 1997 2016 TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49399812 1997 2016 TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 1997 2016TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 1998 2017CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 1998 2017CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 1998 2017CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 1999 2018CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 1999 2018CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 1999 2018CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 2000 2019TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 2000 2019TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 2000 2019TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 2001 2020GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 2001 2020GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 2001 2020GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 2002 2021GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 2002 2021GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 2002 2021GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 2003 2022TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 2003 2022TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 2003 2022TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 2004 2023CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 2004 2023CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 2004 2023CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 2005 2024GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 2005 2024GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 2005 2024GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 2006 2025CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 2011 2030GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 2013 2032CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 2016 2035GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 2016 2035GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 2061 2080ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 395181 2073 2092 AAACAGCTGCCAACCTGCCC5-10-5 87.5 52 405975 2075 2094 CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361405976 2080 2099 TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 2082 2101AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 399992 2095 2114GTGCTGACCACACAGTCCTG 3-14-3 92.6 130 405978 2105 2124GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 2108 2127GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 2110 2129GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 2112 2131CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 2114 2133ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 2116 2135CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 2118 2137GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 2120 2139GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 2168 2187CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 2170 2189AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 2175 2194GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 2177 2196CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 410776 2245 2264GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 2265 2284TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 2295 2314CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 2355 2374CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 2355 2374CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 2355 2374CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 2356 2375ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 2356 2375ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 2356 2375ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 2357 2376GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 2357 2376GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 2357 2376GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 2358 2377TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 2358 2377TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 2358 2377TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 2359 2378TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 2359 2378TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 2359 2378TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 2360 2379CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 2360 2379CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 2360 2379CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 2362 2381TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 2375 2394CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410780 2405 2424GCAGGCCACGGTCACCTGCT 5-10-5 60.9 417 410781 2442 2461GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 2560 2579CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 2560 2579CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 2560 2579CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 2561 2580GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 2561 2580GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 2561 2580GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 2562 2581AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 2562 2581AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 2562 2581AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 2563 2582CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 2563 2582CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 2563 2582CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 2564 2583GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 2564 2583GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 2564 2583GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 2565 2584GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 2565 2584GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 2565 2584GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 2566 2585CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 2566 2585CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410570 2566 2585CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 2567 2586CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 2567 2586CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 2567 2586CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 2568 2587TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 2568 2587TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 2568 2587TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 405988 2568 2587TCCGGCAGCAGATGGCAACG 5-10-5 84.5 426 410572 2569 2588CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 2569 2588CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 2569 2588CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 2570 2589GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 2570 2589GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 2570 2589GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 2580 2599CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 2590 2609GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 2764 2783CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 2852 2871ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 3083 3102ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 3084 3103GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 3085 3104AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 3086 3105GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 3087 3106GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 3087 3106GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 3088 3107TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 3089 3108CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 3090 3109GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 3091 3110AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 3157 3176GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 3158 3177GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 3159 3178TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 3160 3179CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 3161 3180TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 3162 3181CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 3163 3182CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 3164 3183GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 3165 3184AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 3243 3262AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition ofPCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited atleast 80% inhibition of PCSK9 mRNA levels: 395163, 395165, 395166,395168, 395181, 395186, 395187, 395194, 399798, 399801, 399804, 399806,399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399912,399916, 399935, 399936, 399954, 399960, 399962, 399967, 399968, 399969,399973, 399978, 399992, 405869, 405870, 405871, 405872, 405873, 405874,405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883,405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897,405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906,405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916,405920, 405921, 405922, 405940, 405941, 405943, 405944, 405949, 405952,405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973,405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987,405988, 405996, 405997, 406026, 406028, 406029, 406030, 406031, 406032,406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043,406044, 406045, 410562, 410583, 410589, 410590, 410591, 410592, 410593,410595, 410633, 410634, 410658, 410664, 410666, 410667, 410668, 410671,410677, 410713, 410714, 410715, 410716, 410723, 410724, 410754, 410757,410759, 410760, and 410769. The target segments to which these antisenseoligonucleotides are targeted are active target segments. The targetregions to which these antisense oligonucleotides are targeted areactive target regions.

ISIS Nos 395165, 395166, 395168, 395181, 395186, 395187, 395194, 399801,399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916,399936, 399954, 399962, 399969, 399978, 399992, 405869, 405871, 405872,405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882,405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897,405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907,405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922,405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961,405974, 405977, 405978, 405979, 405980, 405982, 405987, 405996, 405997,406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039,406040, 406041, 406042, 406043, 406044, 406045, 410562, 410583, 410591,410592, 410593, 410668, 410677, 410716, 410724, 410757, and 410760 eachexhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 395168, 395186, 395187, 395194, 399801, 399817,399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916, 399954,399969, 399978, 399992, 405873, 405874, 405875, 405876, 405878, 405879,405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895,405899, 405903, 405904, 405905, 405906, 405907, 405910, 405913, 405914,405916, 405941, 405953, 405955, 405956, 405959, 405960, 405978, 405996,405997, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041,406042, 406043, 406044, 410592, 410668, and 410760 each exhibited atleast 90% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978,405997, 406030, 406040, 406041, 406044, and 410760 each exhibited atleast 95% inhibition of PCSK9 mRNA levels.

Example 4: Antisense Reduction of Human PCSK9 mRNA in HepG2 Cells: DoseResponse Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at variousdoses in HepG2 cells. Cells were plated at densities of 10,000 cells perwell and treated with nM concentrations of antisense oligonucleotide asindicated in Tables 20 and 21. After a treatment period of approximately24 hours, RNA was isolated from the cells and PCSK9 mRNA levels weremeasured by quantitative real-time PCR, as described herein. Twodifferent human PCSK9 primer probe sets were used to measure mRNAlevels. Results with Primer Probe Set 2740 are shown in Table 20, andmRNA levels measured using primer probe set 2823 are shown in Table 21.Results are presented as percent PCSK9 mRNA levels relative to acontrol, as adjusted according to total RNA content measured byRIBOGREEN®. As illustrated in Tables 20 and 21, PCSK9 mRNA levels werereduced in a dose-dependent manner.

TABLE 20 Antisense Reduction of PCSK9 mRNA in HepG2 cells, Primer ProbeSet 2740 6.25 12.5 Isis No nM nM 25.0 nM 50.0 nM 100.0 nM 200.0 nM399819 93 113 84 71 26 12 395185 81 64 50 35 24 13 399916 65 56 41 18 2025 399907 68 66 42 26 30 25 399954 71 63 38 22 16 15 395165 71 67 47 2921 10 399936 65 51 31 20 30 28 399793 61 57 44 31 39 15 399969 57 54 4027 25 19 395152 75 58 58 49 24 28

TABLE 21 Antisense Reduction of PCSK9 mRNA in HepG2 cells, Primer ProbeSet 2823 6.25 12.5 Isis No nM nM 25.0 nM 50.0 nM 100.0 nM 200.0 nM399819 80 264 87 85 34 15 395185 86 66 57 45 29 22 399916 75 47 45 20 3045 399907 67 72 47 29 31 32 399954 59 50 35 17 22 27 395165 74 54 41 3726 22 399936 62 41 33 32 28 42 399793 62 51 38 36 44 27 399969 73 40 5828 29 35 395152 78 53 64 49 22 35

Example 5: Antisense Reduction of Human PCSK9 mRNA in Hep3B Cells: DoseResponse Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at variousdoses in Hep3B cells. Cells were plated at densities of 4,500 cells perwell and treated with nM concentrations of antisense oligonucleotide asindicated in Table 22. ISIS 141923 is not complementary to any knowngene sequence and is used in this and the following experiments as anegative control. After a treatment period of approximately 24 hours,RNA was isolated from the cells and PCSK9 mRNA levels were measured byquantitative real-time PCR, as described herein. Two different humanPCSK9 primer probe sets were used to measure mRNA levels. Results withPrimer Probe Set 2740 are shown in Table 22. Results are presented aspercent PCSK9 mRNA levels relative to a control, as adjusted accordingto total RNA content measured by RIBOGREEN®. As illustrated in Table 22,PCSK9 mRNA levels were reduced in a dose-dependent manner.

TABLE 22 Antisense Reduction of PCSK9 mRNA in Hep3B cells, Primer ProbeSet 2740 Isis No. 2.78 nM 8.33 nM 25.0 nM 75.0 nM 399819 127.5 85.0 36.121.9 405891 133.7 111.4 40.2 27.2 406008 144.6 108.3 49.9 17.7 395186124.4 106.4 42.2 30.7 395185 150.7 106.6 53.6 38.1 405994 141.9 101.249.3 30.8 405988 148.1 117.4 47.8 25.2 406033 138.1 109.9 62.4 35.3395187 133.0 115.6 59.9 31.6 405995 124.1 109.2 57.8 42.9 406023 130.0103.5 67.1 28.0 399900 131.2 95.7 81.1 30.9 301012 121.2 100.1 63.8 49.6395165 113.2 86.9 41.6 13.3 405879 124.6 88.8 44.1 13.1 405991 100.6101.4 95.9 41.0 405923 130.4 115.0 68.9 35.1 395152 143.8 98.6 61.3 59.0405881 100.7 77.4 50.2 4.5 141923 131.4 131.9 144.0 167.3

Example 6: Antisense Reduction of Human PCSK9 mRNA in HeLa Cells: DoseResponse Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at variousdoses in HeLa cells. Cells were plated at densities of 5,000 cells perwell and treated with nM concentrations of antisense oligonucleotide asindicated in Table 23. After a treatment period of approximately 24hours, RNA was isolated from the cells and PCSK9 mRNA levels weremeasured by quantitative real-time PCR, as described herein. Twodifferent human PCSK9 primer probe sets were used to measure mRNAlevels. Results with Primer Probe Set 2740 are shown in Table 23.Results are presented as percent PCSK9 mRNA levels relative to acontrol, as adjusted according to total RNA content measured byRIBOGREEN®. As illustrated in Table 23, PCSK9 mRNA levels were reducedin a dose-dependent manner.

TABLE 23 Antisense Reduction of PCSK9 mRNA in HeLa cells, Primer ProbeSet 2740 Isis No. 2.963 nM 8.8889 nM 26.6667 nM 80.0 nM 399819 57 32 1614 405891 57 46 17 18 406008 45 27 11 5.3 395186 40 28 11 6.6 395185 5332 21 17 405994 48 34 20 13 405988 44 32 12 7.3 406033 61 42 25 12395187 42 35 12 6.6 405995 54 39 21 15 406023 76 46 25 6.2 399900 85 4823 6.4 301012 121 109 76 65 395165 61 30 10 2 405879 64 32 9.7 1 40599164 35 24 10 405923 71 45 22 5 395152 73 34 13 3.1 405881 58 33 19 2.8141923 122 118 131 140

Example 7: Antisense Reduction of Human PCSK9 mRNA in PrimaryHepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisenseinhibition of PCSK9 in human primary hepatocytes. Human primaryhepatocytes were purchased from a commercial supplier, and culturedaccording to routine culture procedures. Cells were treated with 10, 25,50, 150, or 300 nM of antisense oligonucleotide for a period of 24hours, after which RNA was isolated and PCSK9 mRNA was measured byreal-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes wereIsis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954,399978, and 399992. The antisense oligonucleotides inhibited PCSK9 in adose-dependent manner in human primary hepatocytes.

Monkey primary hepatocytes were similarly treated with 10, 25, 50, 150,or 300 nM concentrations of antisense oligonucleotide targeted to PCSK9.The antisense oligonucleotides tested in monkey primary hepatocytes wereIsis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954,399978, and 399992. Each of these antisense oligonucleotides is fullycomplementary to a monkey PCSK9 nucleic acid. The antisenseoligonucleotides inhibited PCSK9 in a dose-dependent manner in monkeyprimary hepatocytes.

Example 8: Additional Antisense Reduction of Human PCSK9 mRNA in PrimaryHepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisenseinhibition of PCSK9 in human primary hepatocytes. Human primaryhepatocytes were purchased from a commercial supplier, and culturedaccording to routine culture procedures. Cells were treated with 10, 25,50, 150, or 300 nM of antisense oligonucleotide for a period of 24hours, after which RNA was isolated and PCSK9 mRNA was measured byreal-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes wereIsis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988,405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in adose-dependent manner in human primary hepatocytes as shown in Table 24.

TABLE 24 Dose-dependent Reduction in Human PCSK9 mRNA Expression inHuman Primary Hepatocytes ISIS No. 10 nM 25 nM 50 nM 150 nM 300 nM395165 76 48.7 22.5 20.2 16.2 395185 74.5 42.9 52.6 25.5 15.7 395186123.8 58.3 34.4 21 19.9 395187 98.7 43.5 29.7 17.4 21.1 405879 119.2176.6 64.9 48.4 47.7 405881 222.4 115.3 49.5 22.3 79.3 405891 169.9138.7 73.9 86.9 48.7 405988 85.2 105.4 67.8 43.9 43.8 405994 95.7 83.343.5 24.6 10.5 406008 155 101.7 116 41.7 32.4

Example 9: Antisense Reduction of PCSK9 mRNA in Cyno Primary Hepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisenseinhibition of PCSK9 in cyno primary hepatocytes. Cyno primaryhepatocytes were purchased from a commercial supplier, and culturedaccording to routine culture procedures. Cells were treated with 10, 25,50, 150, or 300 nM of antisense oligonucleotide for a period of 24hours, after which RNA was isolated and PCSK9 mRNA was measured byreal-time PCR as described herein.

The antisense oligonucleotides tested in cyno primary hepatocytes wereIsis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988,405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in adose-dependent manner in cyno primary hepatocytes as shown in Table 25.

TABLE 25 Dose-dependent PCSK9 mRNA Inhibition in Cyno PrimaryHepatocytes ISIS No. 10 nM 25 nM 50 nM 150 nM 300 nM 395165 36.3 29.914.9 8.6 3.2 395185 65.0 31.9 35.1 16.6 12.0 395186 65.8 42.3 21.6 23.316.7 395187 61.3 32.9 13.2 7.7 9.02 405879 24.3 21.9 7.7 7.1 3.0 40588175.0 33.5 26.5 11.2 4.9 405891 52.4 24.8 11.2 10.8 6.7 405988 59.4 23.722.5 10.7 8.2 405994 64.8 52.7 22.2 7.9 6.5 406008 56.3 28.2 26.5 13.413.3

Example 10: Antisense Reduction of Mouse PCSK9 mRNA In Vitro

Antisense oligonucleotides were designed to target murine PCSK9, andwere evaluated for their ability to reduce PCSK9 mRNA in primary mousehepatocytes.

Primary mouse hepatocytes were treated with antisense oligonucleotidesat doses of 50, 150, or 300 nM, for a period of 24 hours. RNA wasisolated using QIAGEN® RNeasy isolation kits and subjected toquantitative real-time PCR using commercially available reagents(Invitrogen, Carlsbad, Calif.). PCSK9 mRNA levels were measured using amouse PCSK9 primer probe set, and normalized to G3PDH mRNA levels and/orRIBOGREEN® levels.

ISIS 394814 (GAGCAACTTCGGAGGCAGC, SEQ ID NO: 456) was identified as apotent inhibitor of mouse PCSK9. PCSK9 mRNA levels ISIS 394814 yielded a50% reduction in PCSK9 mRNA at a concentration of 25 nM (i.e., an IC50of 25 nM) after a 24 hour incubation with primary mouse hepatocytes. Noeffect on cultured cell viability was observed.

Example 11: Antisense Reduction of PCSK9 mRNA in an Animal Model ofHyperlipidemia: High Fat Fed Mice Treatment

C57BL/6 mice fed a high-fat diet are routinely employed as an animalmodel of hyperlipidemia, as well as an animal model of atherosclerosis.Accordingly, to evaluate the effects of PCSK9 antisense inhibition onserum lipid levels in vivo, ISIS 394814 was evaluated in C57BL/6 micefed a high-fat diet. C57BL/6 mice 4-5 weeks of age were obtained fromthe Jackson Laboratory and maintained on a diet consisting of 60% fat.Treatment groups of 5 mice each were as follows: a group treated withISIS 394814; a control group treated with saline; and control grouptreated with an oligonucleotide not complementary to any known genesequence (ISIS 141923). Oligonucleotide or saline was administeredintraperitoneally twice weekly, for a period of 6 weeks; oligonucleotidedoses were 50 mg/kg. After the treatment period, whole blood wascollected for analysis of serum parameters, and whole liver wascollected for RNA analysis, protein analysis, and histologicalevaluation. Statistical analyses included a nonparametric, two-tailedt-test comparison of experimental samples (ISIS 394814) to controlsamples (saline or control oligonucleotide).

RNA Analysis

Liver RNA was isolated for real-time PCR analysis of PCSK9, LDL-receptorand apolipoprotein B mRNA levels. Treatment with ISIS 394814 resulted ina 92% reduction in PCSK9 mRNA levels, while no significant reduction inPCSK9 mRNA levels was observed in ISIS 141923-treated or saline-treatedmice. Antisense inhibition of PCSK9 expression did not significantlyaffect liver LDL-receptor or liver apolipoprotein B mRNA levels.

Protein Analysis

Immunoblotting was performed to assess the effects of PCSK9 antisenseinhibition on LDL-receptor protein and apolipoprotein B levels. Proteinisolated from mouse liver was subjected to electrophoresis, transferredto a polyvinylidene-fluoride membrane, and subsequently probed with ananti-mouse LDL-receptor antibody and a scavenger receptor B1 (SR-B1)antibody. Mouse plasma was similarly subjected to electrophoresis andimmunoblotting using an anti-mouse apolipoprotein B antibody. Secondaryantibodies conjugated to peroxidase were used to detect primaryantibodies. Protein bands were visualized using the ECL plus Westernblot detection kit (Amersham Biosciences, UK) and quantified usingImageQuant™ analysis software (MolecularDynamics, Santa Clara, Calif.).

While LDL-receptor mRNA levels were not significantly affected,antisense inhibition of PCSK9 resulted in an approximate 2-fold increasein the level of hepatic LDL-receptor protein relative to 141923-treatedcontrols. No changes in SR-B1 protein were observed. Furthermore, serumapoB-100 levels were significantly reduced by 50%, relative to141923-treated controls. Serum apoB-48 levels were significantlyincreased by approximately 3-fold relative to 141923-treated controls.No significant changes in apoA-I protein were observed, relative to141923-treated controls.

The mRNA levels of the RNA editing enzyme apobec-1 were also measured.Apobec-1 is responsible for the RNA editing that produces apoB-48 inmurine liver and intestine. Human apobec-1 is expressed only inintestinal cells, thus these cells are the source of apoB-48 in humans.Antisense inhibition of PCSK9 resulted in an increase in murine hepaticapobec-1 mRNA levels by approximately 2.7 fold compared tosaline-treated controls

Serum Lipid Analysis

Plasma concentrations of total cholesterol, LDL-C, HDL-C, freecholesterol, triglycerides, glucose, ketones, transaminases, andphospholipids were measured using an automated clinical chemistryanalyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein andcholesterol profiling was performed as described by Crooke et al. (LipidRes., 2005, 46, 872-884) using a Beckman System Gold 126 HPLC system, a507e refrigerated autosampler, a 126 photodiode array detector (BeckmanInstruments, Fullerton, Calif.), and a Superose 6 HR 10/30 column(Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions were measured at awavelength of 505 nm and validated with a cholesterol calibration kit.(Sigma).

Administration of ISIS 394814 resulted in a 52% reduction in totalcholesterol (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS394814, 87 mg/dL±19, p<0.005) and a 36% reduction in LDL-C (saline 22mg/dL±4; ISIS 141923, 25 mg/dL±2; ISIS 394814, 14 mg/dL±2, p<0.005).Serum free cholesterol was reduced by 25% (saline 41 mg/dL±10; ISIS141923, 58 mg/dL±3; ISIS 394814, 31 mg/dL±5, p<0.005) and phospholipidswere reduced 54% (saline, 354 mg/dL±29; ISIS 141923, 383 mg/dL±21; ISIS394814, 169 mg/dL±35, p<0.0001). HDL-C was also reduced by approximately54% (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS 394814, 87mg/dL±19; p<0.0001). HPLC profiling confirmed the reduction of LDL andHDL lipid classes.

Liver Triglyceride Analysis

Liver triglyceride content was measured according to routineexperimental procedures, for example, per procedures described by Desaiet al., Diabetes, 2001, 50:2287-2295.

Antisense inhibition of PCSK9 for 6 weeks reduced liver triglyceridecontent by approximately 65% (p=0.01) relative to saline controls. Nostatistically significant changes in liver triglyceride contentfollowing 141923 treatment were observed.

Accordingly, one embodiment is a method of lowering LDL-C levels throughthe administration of an antisense oligonucleotide targeted to a PCSK9nucleic acid. An additional embodiment includes a method of loweringtotal cholesterol through the administration of an antisenseoligonucleotide targeted to a PCSK9 nucleic acid. An addition embodimentis lowering liver triglycerides by administering an antisenseoligonucleotide targeted to a PCSK9 target nucleic acid.

Example 12: Antisense Reduction of PCSK9 mRNA in LDL-RDeficient/apoB-100 Animals

LDL-receptor (LDL-R)-deficient/apoB-100 animals do not express the LDL-Rgene, and express only the apoB-100 form of apoB. PCSK9 antisenseoligonucleotide was administered to LDL-R deficient/apoB-100 mice, toevaluate the effects of antisense inhibition of PCSK9 in the absence ofthe LDL-R. Mice 4-5 weeks of age were maintained on a standard mousediet. Treatment groups of 5 mice each were as follows: a group treatedwith ISIS 394814; a control group treated with saline; and control grouptreated with an oligonucleotide not complementary to any known genesequence (ISIS 141923). Oligonucleotide or saline was administeredintraperitoneally twice weekly, for a period of 6 weeks; oligonucleotidedoses were 50 mg/kg (100 mg/kg/wk total). Real-time PCR of liver mRNAand serum analyses were performed as described for the C57Bl/6 study.

Antisense inhibition of PCSK9 reduced liver PCSK9 mRNA by approximately90%, relative to saline controls. However, no reduction in serumcholesterol was observed. Liver triglyceride levels were unaffected byantisense inhibition of PCKS9 in these mice. An 80% increase in liverapobec1 mRNA levels was observed, relative to saline controls. Thesedata suggest that a functioning LDL-R is required for cholesterol andliver triglyceride reduction via PCSK9 antisense inhibition.

Example 13: Antisense Reduction of PCSK9 mRNA in Mouse PrimaryHepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisenseinhibition of PCSK9 in mouse primary hepatocytes. Mouse primaryhepatocytes were purchased from a commercial supplier, and culturedaccording to routine culture procedures. Cells were treated with 6.25,12.5, 25, 50, 100, or 200 nM of antisense oligonucleotide for a periodof 24 hours, after which RNA was isolated and PCSK9 mRNA was measured byreal-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes wereIsis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988,405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in adose-dependent manner in human primary hepatocytes as shown in Table 26.

TABLE 26 Dose-Dependent Reduction PCSK9 mRNA in Mouse PrimaryHepatocytes 6.25 12.5 ISIS No nM nM 25.0 nM 50.0 nM 100.0 nM 200.0 nM395165 95.2 94.9 81.7 66.3 57.5 29.9 395185 109.2 100 90.5 81.2 64.735.8 395186 101.9 91.5 77.8 59.1 39.5 13.7 395187 102.4 96.1 83.9 66.440 15.3 405879 107.3 97.5 87.8 76.6 63 43.1 405881 104.8 102.1 98.1 84.968.7 48.9 405891 104.9 103.3 101.5 88.9 79.6 49.5 405988 97.3 101.1 91.782.3 61.2 34.8 405994 110.2 101 111.6 94.4 80.1 57.7 406008 99.5 91.383.9 73.7 61.8 39.4 157700 97.8 98.3 96.2 94.3 83.2 76.4 141923 84.3 8079.9 77.3 98.8 64.2

Example 14: Antisense Reduction of Human PCSK9 mRNA in Hep3B and HeLaCells: Dose Response Experiment

A subset of the antisense oligonucleotides that inhibited PCSK9expression at high levels in the cell culture systems described abovewas selected for further screening. The group of selected antisensecompounds is shown in Table 27. Table 27 shows the Isis number,nucleotide sequence, and percent inhibition obtained in Hep3B cellsusing the protocol disclosed in Example 3 (see Table 17) for the variousantisense compounds.

TABLE 27 SEQ % ID Isis No. Sequence 5′-3′ Motif inhibition NO 405881CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 399819 CCCACTCAAGGGCCAGGCCA 5-10-593.4 65 395165 ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 405879CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 406008 CTTGGTGAGGTATCCCCGGC 5-10-595.2 188 405891 TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 395186CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 405988 GGCAGCACCTGGCAATGGCG 5-10-595.9 381 405994 GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 406023TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 395187 GGCAGCAGATGGCAACGGCT 5-10-592.7 62 395185 CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 406033CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 405923 GGCATTGGTGGCCCCAACTG 5-10-594.3 288 399900 GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 405995TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 405991 GACACGGGTCCCCATGCTGG 5-10-591.9 394 406005 CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 399793CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 395152 ACGCAAGGCTAGCACCAGCT 5-10-593.3 7

The IC₅₀ of the various antisense compounds was determined usingdifferent cell culture systems. Antisense oligonucleotides targeted toPCSK9 were tested at various doses in Hep3B or HeLa cells, for example.Cells were plated at densities of 4,500 cells or 5,000 per well,respectively, and treated with of antisense oligonucleotides at variousconcentrations. After a treatment period of approximately 24 hours, RNAwas isolated from the cells and PCSK9 mRNA levels were measured byquantitative real-time PCR, as described herein, relative to untreatedcontrol cells. Two different human PCSK9 primer probe sets were used tomeasure mRNA levels. Results with Primer Probe Set 2740 are shown inTable 28. PCSK9 mRNA levels were adjusted according to total RNA contentas measured by RIBOGREEN®. Table 28 shows IC50 values (nM) obtained fromtwo experiments with Hep3B cells and from one experiment with HeLacells.

TABLE 28 IC₅₀ values Hep3B Hep3B HeLa SEQ IC₅₀ IC₅₀ IC₅₀ ID ISIS # (nM)(nM) (nM) PCSK9 Target Sequence NO 405881 21 26 13 CACCCTTGGCCACGCCGGCA250 399819 21 28 12 CCCACTCAAGGGCCAGGCCA 65 395165 23 28 13ACCCTTGGCCACGCCGGCAT 28 405879 24 28 14 CCTTGGCCACGCCGGCATCC 248 40600831 35 8 CTTGGTGAGGTATCCCCGGC 188 405891 32 38 14 TCCTCAGGGAACCAGGCCTC352 395186 33 33 7 CTTTGCATTCCAGACCTGGG 60 405988 34 42 8GGCAGCACCTGGCAATGGCG 381 405994 35 39 9 GCAGTGGACACGGGTCCCCA 400 40602340 50 18 TGGTATTCATCCGCCCGGTA 212 395187 41 44 8 GGCAGCAGATGGCAACGGCT 62395185 42 43 11 CACGGGTCCCCATGCTGGCC 59 406033 44 61 15CCTGCCAGGTGGGTGCCATG 237 405923 47 41 20 GGCATTGGTGGCCCCAACTG 288 39990048 44 21 GGTCCTCAGGGAACCAGGCC 50 405995 48 63 12 TGGCAGTGGACACGGGTCCC402 405991 63 52 15 GACACGGGTCCCCATGCTGG 394 406005 63CGGGCAGTGCGCTCTGACTG 180 399793 >75 CCTCGGAACGCAAGGCTAGC 8 395152 46 16ACGCAAGGCTAGCACCAGCT 7

Several of the antisense compounds examined above were selected forfurther study. These antisense compounds exhibited at least about 93%inhibition of human PCSK9 mRNA expression in the Hep3B cell culturesystem, as shown in Tables 17-19, for example. As shown in Table 28, theselected antisense compounds also displayed IC50 parameters of less thanabout 45 nM when tested with Hep3B cells and less than about 15 nM whentested with HeLa cells. The selected antisense compounds, listed inTable 29, are Isis 405881, Isis 395165, Isis 405879, Isis 406008, Isis405891, Isis 395186, Isis 405988, Isis 405994, Isis 395187, and Isis395185. Isis 405879, for example, showed superior results in both Hep3Band HeLa cells, with IC₅₀ values of about 26 nM and 14 nM, respectively.

Example 15: Dose-Dependent Antisense Reduction of PCSK9 mRNA in HumanPrimary Hepatocytes; Cynomolgus Monkey (“Cyno”) Primary Hepatocytes; andHuman HEK-293 Cells

The selected antisense oligonucleotides were further tested for dosedependent antisense inhibition in human primary hepatocytes, cynoprimary hepatocytes, and HEK-293 cells expressing human PCSK9. Cellswere purchased from commercial suppliers and cultured according toroutine culture procedures. Cells were treated with 10, 25, 50, 150, or300 nM antisense oligonucleotide (i.e., a five-point dose response test)for a period of 24 hours, after which RNA was isolated. PCSK9 mRNA wasmeasured by real-time PCR, as described herein.

Results of the five-point dose response tests using HEK-293 cells areshown in FIG. 2. Results of the five-point dose response tests usingcyno primary hepatocytes are shown in FIG. 3. The IC₅₀ results,expressed in nM units, for the five-point dose response test using humanprimary hepatocytes, cyno primary hepatocytes, HEK-293 cells, Hep3Bcells, and HeLa cells are summarized in Table 29.

TABLE 29 IC₅₀ (nM) for Selected Antisense Compounds Isis No. 395165395185 395186 395187 405879 405881 405891 405988 405994 406008 SEQ IDNO: 28 59 60 62 248 250 352 381 400 188 Human Primary 24 33 39 30 ND 50297 166 56 163 Hepatocytes Cyno Primary 5 17 18 14 2 19 10 12 21 11Hepatocytes HEK-293 40 116 6 11 35 86 21 11 17 27 Hep3B 28 43 33 44 2826 38 42 39 35 HeLa 13 11 7 8 14 13 14 8 9 8

ISIS 405879 was tested at various doses in human primary hepatocytes.RNA was isolated from the cells and PCSK9 mRNA levels were measured byquantitative real-time PCR, as described herein. Human PCSK9 primerprobe set 2740 was used to measure mRNA levels. Results are shown inTable 29.1.

TABLE 29.1 Dose-dependent Reduction in Human PCSK9 mRNA Expression inHuman Primary Hepatocytes ISIS 300 NO. nM 200 nM 150 nM 100 nM 50 nM 25nM 10 nM 405879 13 18 26 30 27 54 75

Isis 405879 has an IC50 value of 29 nM in human primary hepatocytes.Overall Isis 405879 displays superior activity, particularly in cynoprimary hepatocytes. Isis 405879 was thus further tested in the variouscell models, and the results of the additional testing are shown inTable 30. In these experiments, Isis 405879 was administered to thecells using lipofectin transfection reagents. PCSK9 mRNA was assayed byPCR following 24 hr incubation after transfection. These results confirmthat Isis 405879 potently inhibits PCSK9 mRNA expression at lowconcentrations.

TABLE 30 Isis 405879 activity in cell culture models HepatocyteTransgenic Hep3B HepG2 HeLa Transgenic Primary Primary HEK293 CellsCells Cells Mouse Hepatocyte Hepatocyte cells (human) (human) (human)(h-PCSK9) (human) (cyno) (h-PCSK9) Mean 17 5 14 140 22 7.4 18 IC₅₀ (nM)

These experiments confirm that the presently disclosed antisenseoligonucleotides are capable of potently inhibiting PCSK9 mRNAexpression in a variety of cell model systems, including a human primarycell line. Exemplary antisense compounds inhibit PCSK9 mRNA expressionwith IC50 values in the nM range.

The transgenic PCSK9 cDNA construct expressed in mouse hepatocytes isthought to be rapidly exported to the cytoplasm, because it does notrequire post-transcriptional editing. The antisense oligonucleotidesprovided herein act by recruiting the RNase H enzyme, which isrestricted to the nucleus. Therefore, inhibition by antisenseoligonucleotides in this cell type is significantly diminished.

Example 16: In Vivo Testing Procedures 16.1. High Fat (HF)-FedHyperlipidemic Mouse Model

Antisense compounds may be tested in a high fat-fed, hyperlipidemicmouse model to test efficacy of the antisense compounds in reducingPCSK9 mRNA expression, increasing LDL-R expression, reducing totalcholesterol and LDL, and affecting other metrics of antisense compoundefficacy. A representative hyperlipidemic mouse model is described inGraham et al., J. Lipid Res. (2007) 48: 763-767.

All animal experiments were conducted in accordance with InstitutionalAmerican Association for the Accreditation of Laboratory Animal Careguidelines. C57BL/6 mice were obtained from Jackson Laboratory(http://www.jax.org). A majority of the mice were male, and studies wereinitiated when animals were 4-5 weeks of age. The mice were maintainedon a 12 h light/12 h dark cycle and fed ad libitum. C57BL/6 mice werefed a diet consisting of 60% lard (Research Diets, New Brunswick, N.J.),whereas experimental Ldlr-deficient/apoB-100 mice were fed regular chow.

Antisense compounds were administered twice weekly (50 mg/kg) for 6weeks by intraperitoneal injection (10 mg/mL dosing solution formulatedin saline; saline control). After the treatment period, whole blood wascollected for analysis of serum parameters, and whole liver wascollected for RNA analysis, protein analysis, and histologicalevaluation.

16.2. H-ApoB/CETP Transgenic Mouse Model

A transgenic mouse model that expresses both human apolipoprotein (apo)B and human cholesteryl ester transfer protein (CETP) provides anotheruseful animal model system to test the efficacy of the presentlydisclosed antisense compounds. The lipoprotein cholesterol distributionin the serum of a chow-fed h-apoB/CETP mouse resembles a human profile.Specifically, the percentages of total cholesterol within the HDL, LDL,and VLDL fractions of apoB/CETP animals are approximately 30%, 65%, and5%, respectively, similar to the distribution of cholesterol in theplasma of normolipidemic humans. The h-apoB/CETP mouse model isdescribed further in Grass et al., J. Lipid Res. (1995) 36: 1082-1091.

16.3. H-PCSK9 Transgenic Mouse Model

A transgenic mouse expressing human PCSK9 provides another useful animalmodel to study in vivo repression of PCSK9 mRNA by the present antisensecompounds. H-PCSK9 transgenic mice overexpress human PCSK9 in liver andsecrete large amounts of the protein into plasma, which increases plasmaLDL-C concentrations to levels similar to those of LDLR-knockout mice.H-PCSK9 transgenic mice were constructed by inserting a cDNA encodinghuman PCSK9 into a pLiv-11 vector that contained the constitutive humanapoE promoter and its hepatic control region, as described in Simonet etal., J. Biol. Chem. (1993) 268: 8221-8229. Transgenic mice weregenerated by injecting linearized pLiv-11-hPCSK9 into the fertilizedeggs as described in Shimano et al., J. Clin. Invest. (1996) 98:1575-1584. H-PCSK9 transgenic mice are further described in Lagace etal., J. Clin. Invest. (2006) 116: 2995-3005.

16.4. Mouse and Rat Models for In Vivo Tolerability, Half-Life andTissue Distribution

Mouse (CD1) and rat (Sprague-Dawley) models were used to test the invivo tolerability of various antisense compounds. In the mouse CD1model, antisense compounds were overdosed at 100 mg/kg/wk for six weeks.Possible proinflammatory effects and hepatotoxcity were assayedfollowing administration of the antisense compounds. Serum levels ofaspartate aminotransferase (AST), alanine aminotransferase (ALT), bloodurea nitrogen (BUN), creatinine, albumin, and bilirubin were assayed.Weights of livers, kidneys, and spleens of animals were determined.Histopathologic screening was conducted to detect hepatocellularswelling, multifocal apoptosis, inflammatory cell infiltrates in theliver or lungs, or follicular hyperplasia. Additional studies wereconducted in the rat model to screen for renal toxicity or high renalaccumulation and proinflammatroy effects. In these experiments,antisense compounds were administered at 60 or 100 mg/kg/wk for sixweeks. In addition to the markers examined above, the urineprotein/creatine ratio and proteinuria were assayed.

The half-life of administered antisense compounds also was examined inthe target organ, i.e., liver, in the mouse model. For theseexperiments, antisense compounds were administered twice a week for twoweeks at 50 mg/kg, and target organs were then harvested and assayed forthe presence of the antisense compounds, as described below.

The distribution of administered antisense compounds in various organsalso was examined in the rat model. Rats in these studies wereadministered 20 mg/kg antisense compounds twice weekly for three weeks.Livers and kidneys of test animals were then harvested and assayed forthe presence of the antisense compounds, as described below.

16.5. Cynomolgus Monkey (M. fascicularis) Model

Potencies and pharmacodynamics of antisense compounds were compared in acyno model to identify the compounds exhibiting the best combination ofhigh potency and low renal accumulation, when given subcutaneously toanimals for a 5-week period. In another study, dosing was conducted overa 13-week period. The cyno monkey is widely used in toxicity testing,and a substantial historic toxicology database is available for thismodel.

The PCSK9 target sequence for an antisense compound may differ in humansversus cyno monkey. As a result, an antisense compound for use in humansmay not be fully complementary to a monkey PCSK9 nucleic acid. Antisensecompounds having identical target sequences in the two species arepreferred. Isis 405881 and Isis 395165 each have a one nucleotidemismatch compared to the cyno sequence, meaning that these antisensecompounds are less preferred in the cyno model.

Cyno monkeys (Charles River BRF, Inc.) were approximately 26 to 32months old and 2.5 to 4 kg in weight on the first day of dosing. Monkeyswere offered water ad libitum and fed a daily ration of 12 biscuitsapproximately 2 hours post dose. Antisense compounds were administeredsubcutaneously at 3 doses every other day during the first week, then byweekly injections. A low dose of 15 mg/kg was used to assess the potencyof the antisense compounds. A high dose of 8 subcutaneous injections of30 mg/kg over 5 weeks or 13 weeks was used to assay renal accumulationand pharmacodynamics. Antisense compounds were administered at aconcentration of 60 or 120 mg/mL, respectively, in a vehicle of sterilephosphate buffered saline. Plasma concentrations of antisense compoundswere determined at each dose level to estimate systemic exposure inmonkeys following subcutaneous administration and to assess whethersystemic exposure was altered after repeat dosing.

To measure the host reaction to the administered antisense compounds, 2mL blood samples were collected from the femoral vein of hostindividuals in tubes of appropriate size containing K₂EDTA. Plasma wasobtained by centrifugation and samples were stored frozen. The analysisof the plasma samples included a determination of the level of plasmaPCSK9 protein. To measure an effect on pharmacodynamic end points, LDL,HDL, and apoB levels also were analyzed. To determine whether the hostindividuals suffered side effects, blood samples were further analyzedfor an effect on the hematological parameters listed in Table 31 and theserum chemistry parameters listed in Table 32:

TABLE 31 Hematology markers erythrocyte count red cell distributionwidth hemoglobin platelet count hematocrit mean platelet volume meancorpuscular volume absolute total and differential leukocyte counts meancorpuscular hemoglobin evaluation of cell morphology concentrationabsolute reticulocyte count

TABLE 32 Serum chemistry markers aspartate aminotransferasetriglycerides alanine aminotransferase glucose gamma glutamyltransferaseurea nitrogen alkaline phosphatase creatinine total bilirubin calciumtotal cholesterol phosphorus total protein sodium albumin potassiumglobulins chloride albumin/globulin ratio

Possible additional side effects were determined by screeningindividuals administered antisense compounds for gross changes in organsor microscopic changes in tissues. A limited necropsy conducted on alltest animals included gross examination of various organs and tissues.Terminal body weights were recorded, and kidneys and spleen were weighedfor all test animals. For microscopic evaluation, liver, kidney, spleenand injection sites from all animals were fixed in 10% neutral bufferedformalin and were processed, embedded in paraffin, sectioned, andstained with hematoxylin and eosin.

Representative tissue samples of liver and kidney cortex from eachscheduled-necropsy monkey from the high-dose and control groups werecollected for ultrastructural examination using electron microscopy.Thin slices (2-4 mm in thickness) of liver and kidney cortex were cutand transferred to a piece of dental wax containing a small volume ofMcDowell-Trump fixative. Thin slices were further minced intoapproximately 1×1-2 mm cubes in McDowell-Trump fixative and analyzed byelectron microscopy.

To evaluate PCSK9 mRNA levels, samples from liver (approximately 1 g)were homogenized using a Polytron tissue disruptor in 10 mL of RLTsolution (Qiagen) and snap-frozen and stored at −70° C. or colder.Additionally, samples (approximately 500 mg each) from kidney werecollected and snap-frozen for mRNA analysis.

Tissue samples from liver and kidney cortex were also analyzed for thepresence of the administered antisense compounds. Antisense compoundspresent in the tissues can be detected and quantified using capillarygel electrophoresis (CGE), as described in Leeds et al, Drug MetabDispos. (1998) 26: 670-675, for example. In this procedure, tissues areweighed, mixed with an internal standard (e.g., a 27-mer (T)oligonucleotide) in proteinase K digestion buffer (2.0 mg/ml proteinaseK in 20 mM Tris-HCl, pH 8.0, 20 mM EDTA, 100 mM NaCl, 0.5% NonidetP-40), and homogenized. Overnight incubation at 37° C. is followed byphenol/chloroform extraction. Tissue extracts are purified by sequentialanion-exchange solid-phase extraction (SPE) followed by reverse-phaseSPE, described in Leeds et al., Anal Biochem (1996) 235: 36-43. Samplesare further desalted by membrane dialysis before analysis usinggel-filled capillaries, as described in Leeds et al. (1996).

Capillary gel electrophoresis separations can be performed as describedby Leeds et al. (1998), using a Beckman P/ACE capillary electrophoresisinstrument (model 5010) with a 27-cm column (effective length, 20 cm)containing 12% polyacrylamide, with 8.3 M urea in 100 mM Tris-borate, pH8.5, as the running buffer. Separation is achieved at 50° C. and 550V/cm. Oligomers eluting from the column are detected by UV absorption at260 nm. Quantification of antisense oligonucleotides is based on thetissue weight extracted and the initial T27 concentration.

Example 17: In Vivo Testing Results

17.1. Reducing PCSK9 mRNA In Vivo Using a Murine Antisense CompoundCorrelates with In Vivo Plasma LDL-C and Liver TG Reduction

In vivo pharmacodynamic studies were performed with the murine antisensecompound Isis 394816 to validate the pharmacodynamic endpoint of PCSK9mRNA reduction. Isis 394816 has the sequence 5′ GGTAAGGTGCGGTAAGTCCT 3′(SEQ ID NO: 462), which targets position 3100 (5′) of the codingsequence of murine PCSK9 mRNA (NCBI Accession No. NM_153565.1; SEQ IDNO: 463). Isis 394816 has a 3-14-3 motif.

Isis 394816 was administered by intraperitoneal injection to anh-apoB/CETP transgenic mouse for 6 weeks at a dosing regimen of 20mg/kg/wk or 50 mg/kg/wk. See Example 16.2. As shown in FIG. 4, Isis394816 administered at either dose caused an approximate 80% reductionin PCSK9 mRNA expression in livers of test animals, compared to controlanimals. As shown in FIG. 5, panel A, Isis 394816 produced a significant(p<0.0001) decrease in the level of plasma LDL-C at the end of sixweeks. Additionally, as shown in FIG. 5, panel B, Isis 394816 produced asignificant (p<0.02) decrease in the level of liver TG at the end of sixweeks.

Additionally, Isis 394816 reduced PCSK9 mRNA expression, plasma LDL-C,and liver TG in a dose dependent manner, as shown in FIG. 6. The potencyof Isis 394816 was much higher in a hyperlipidemic mouse model (seeExample 16.1.) than in a lean mouse model (data not shown). Thereduction of plasma LDL-C positively correlated with the inhibition ofPCSK9 mRNA levels in liver, shown in FIG. 7. These studies demonstratethat the reduction of liver PCSK9 mRNA correlates with the desiredendpoint of reducing plasma LDL-C and liver TG. Accordingly, thereduction of liver PCSK9 mRNA provides a valid pharmacodynamic endpoint.

17.2. Differences in the Half-Life (t_(1/2)) of Antisense Compounds inLiver

The half-lives of selected antisense compounds were assayed in mouse CD1livers, according to the procedures set forth in Example 16.4.Half-lives varied among the tested antisense compounds, which again isattributable to differences in structural features of the antisensecompounds, such as nucleotide sequences and/or motifs. For example, thehalf-life of Isis 405879 (t_(1/2)=27.7 days) was significantly higherthan the half-life of Isis 395165 (20.4 days), although the targetsequences of these two antisense compounds are shifted by only twonucleotides:

Isis 405879: (SEQ ID NO: 248) CCTTGGCCACGCCGGCATCC Isis 395165: (SEQ IDNO: 28) ACCCTTGGCCACGCCGGCATHalf-life data for exemplary antisense compounds is summarized in Table33. Isis 405879 displayed a particularly high half-life in liver.

TABLE 33 Liver t_(1/2) (days) Antisense Compound Liver t_(1/2) (d)395165 20.4 395185 20.2 395186 19.9 395187 13.7 405879 27.7 405881 14.1405891 17.1 405988 14.6 405994 20.2 406008 19.7

17.3. Differences in the Liver and Kidney Distributions of AntisenseCompounds

The antisense compounds listed in Table 29 were administered to rats todetermine the distribution of antisense compounds in liver and kidney.Accumulation of high levels of antisense compounds in the kidneys maycause adverse side effects. Antisense compounds were dosed andadministered as described in Example 16.2. FIG. 8 shows the distributionof the antisense compounds shown in Table 29 in rat liver and kidneys.Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405881, Isis405891, Isis 405988, and Isis 405994 show a particularly advantageousratio of distribution between liver and kidney.

When tissue distribution was tested in a cyno model, further differencesamong antisense compounds were observed. Accumulation of Isis 395186,Isis 405879, and Isis 405891 in liver and kidney was examined afterdosing each antisense compound at two concentrations. See Example 16.5.The results are shown in FIG. 9. Accumulation at each dosingconcentration (15 mg/kg and 30 mg/kg) is shown for kidney (left sidebar) and liver (right side bar). Isis 405879 advantageously accumulatesat a high concentration in liver, even at a dose of 15 mg/kg, andaccumulates at a low level in kidney. Specifically, Isis 405879accumulated in kidneys at a concentration of 1300 μg/g (15 mg/kg dose)to 1800 μg/g (30 mg/kg dose). The average accumulation of antisensecompounds in kidneys is 2500 μg/g (15 mg/kg dose) to 4000 μg/g (30 mg/kgdose). Variation in kidney and liver accumulation is attributable tostructural differences, such as nucleotide sequences and/or motifs,between the antisense compounds.

17.4. Differences in Host Tolerance of Antisense Compounds

Rodent tolerability studies, conducted as described in Example 16.4,showed that antisense compounds were well tolerated. Followingadministration of the exemplary antisense compounds listing in Table 29,serum chemistry markers were normal, with some variably high ALT and ASTlevels observed for Isis 405988 and Isis 405994. Whole body weight andmajor organ weights in CD-1 mice also were not significantly affected byadministration of the exemplary antisense compounds. Histologicalfindings revealed differences between the various antisense compounds,but were consistent with the overall good tolerability for the antisensecompounds. The results of the histological screens are shown in Table34.

17.5. Results of the 5 wk Cyno Toxicology Study

The methods used to conduct the 5 wk cyno toxicology study are describedin Example 16.5. Isis 405879 is referred to as BMS-844421 below and inthe corresponding figures. The PCSK9 mRNA levels assayed by q-RT-PCR inthe cyno liver tissue samples were decreased by BMS-844421 treatment byan average of 65% and 70% (average of males and females) versus salinecontrols at the low and high doses, respectively (FIG. 25). The Effectwas Seen in Both Females and male monkeys, despite relatively highvariability of mRNA level between animals. No dose-dependency was seenbetween the 2 doses. The data are mean+/− for male and female animalscombined.

The levels of BMS-844421 ASO were measured in liver at the end of the 5wk study. As shown in FIG. 26, liver full-length BMS-844421 levelsreached ˜1 mg/gram tissue in both dose groups. This level is consistentwith values from previous experiences with other liver-targeting ASOs.Most of the compound was detected as intact full-length BMS-844421. Nodose-dependency was seen in the ASO liver accumulation data, inagreement with the lack of dose-dependent effects on liver PCSK9 targetmRNA.

Pharmacodynamic responses were assessed in serum lipid assays for thecyno 5 wk ASO study. As seen in FIG. 27, a modest trend (notstatistically significant) toward lowering LDL-C was observed forBMS-844421 treated cyno monkeys at 2.5 and 5 wk of dosing. This trend atboth time points for BMS-844421 relative to baseline was also seen inthe scatterplot of individual animal data in FIG. 28A. HDL/LDL ratioswere sometimes employed as a sensitive indicator of plasma lipid status.A chart of the average ratio of HDL/LDL in the cyno study showed a trendfor increase in the ratio for BMS-844421 treated cynos at 2.5 and 5 wk(FIG. 28B).

Gene expression profiling by Affymetrix microarray was conducted on theRNA samples from this study. The analysis included liver RNA fromanimals administered vehicle alone, BMS-844421 at 15 mg/kg and 30 mg/kgand BMS-844419 and BMS-844423 at 30 mg/kg only. By microarray, therewere significant (p<0.01) transcriptional changes in approximately 1 to2% of genes with approximately 1% expected to change at thissignificance level by chance. For drug-dosed groups relative to controlthere were alterations in inflammation-, immune system-, and lipidhomeostasis-related transcripts with statistically significant animal toanimal variability. Other drug-related hepatic transcriptional changescommon to multiple dose groups were associated with several or a fewgenes in various biological processes and are of unknown significance.

17.6. Results of the 13 wk Cyno Toxicology Study

An additional opportunity to monitor pharmacodynamic responses toBMS-844421 in monkeys was the 13 wk toxicology study, described inExample 16.5. Lean cyno monkeys with low LDL levels were again thesubject for this study, as for the 5 wk cyno monkey study above. At 13wks subcutaneous dosing at 4 different dose levels, BMS-844421suppressed PCSK9 expression as measured by both liver PCSK9 mRNA andplasma PCSK9 protein levels. Despite the inter-animal variability inthese values, it is apparent that BMS-844421 strongly suppressed bothliver mRNA and plasma PCSK9 protein (FIG. 29). 40 mg/kg suppressed PCSK9protein levels very robustly with less variation between animals (FIG.29). No statistically significant differences were seen in plasma LDL-Cor other lipid parameters in this study.

TABLE 34 Histological Findings in CD-1 Mice Antisense Lymph Skeletalcompound Liver Kidney Lung Heart Spleen node Intestine Thyroid SternumMuscle Saline − − − − − − − − − − ISIS 395165 +/++ − −/+ −/+ − − − −ISIS 395185 + − − − −/+ − − − − − ISIS 395186 − − − − − − − − − − ISIS395187 − − − − −/+ − − − − − ISIS 405879 − − − − − − − − − − ISIS 405881inflam − inflam − −/+ −/+ − − − − ISIS 405891 − − − − −/+ −/+ − − − −ISIS 405988 − − − − − − − − − − ISIS 405994 ++ − − − −/+ − − − − − ISIS406008 +/++ − − − − − − − − − Normal: − Minimal changes: −/+ Mildchanges: + Moderate changes: ++ Severe changes: +++

Microscopic evaluation of mouse organs from animals treated with 10different human PCSK-9 antisense compounds revealed that all compoundsgenerally were well tolerated, but that some antisense compounds wereparticularly well tolerated. The following results were seen in thevarious examined organs.

-   -   Liver: Mild (+) to moderate (++) liver injuries (cytoplasmic        swelling to multifocal apoptosis) were seen in Isis 395165        (+/++), Isis 395185 (+), Isis 405988 (+/++), Isis 405994 (++),        and Isis 406008 (+/++) treated livers. Mild inflammatory changes        were seen in Isis 405881 treated livers.    -   Lung: No significant abnormality was seen in treated animals        except for mild histiocytosis noticed in Isis 405881 treated        animals.    -   Spleen: Mild follicular hyperplasia was seen in Isis 395165,        Isis 395185, Isis 395187, Isis 405881, Isis 405891, and Isis        405994 treated spleens.    -   Lymph node: Minimal histiocytosis was observed in Isis 395165,        Isis 405881 and Isis 405891 treated animals.    -   Heart: Minimal levels of macrophage infiltration were seen in        Isis 395165, Isis 395186, Isis 405879, Isis 405881, Isis 405891,        Isis 405988, and Isis 406008 treated hearts.    -   Kidney, Thyroid, small intestine, skeletal muscle and sternum:        No abnormality was visualized.

Differences in the histological findings were seen even between Isis395165 and Isis 405879. As noted above, the target sequences of thesetwo antisense compounds are only shifted by two nucleotide positions.Nevertheless, Isis 405879 advantageously displays no adverse reactions,whereas Isis 395165 elicits a mild inflammatory response in liver,spleen, and lymph nodes. In summary, Isis 405879, Isis 395186, and Isis405988 display superior tolerance, compared to the other antisensecompounds tested in this study.

17.7. Differences in Achieving Pharmacodynamic Endpoints in the CynoModel.

Isis 395186 (“844419”), Isis 405879 (“844421”), and Isis 405891(“844423”) were selected for further study in the cyno model, using theprocedures described in Example 16.6. As set forth above, theseantisense compounds displayed high activity in cell culture models, werewell tolerated, and displayed advantageous tissue distribution andhalf-life properties. Further, none of these antisense compoundstargeted a PCSK9 sequence known to have differences in monkeys andhumans. In addition, the target sequences in human PCSK9 were known notto contain SNPs having a wide distribution in the population.

The antisense compounds were tested for their ability to reduce PCSK9mRNA in cyno liver. See Example 16.6. The results of this experiment areshown in FIG. 10. Despite variability between the tested animals, Isis405879 (labeled “844421”) displayed a superior ability to reduce PCSK9mRNA levels significantly. When administered at a low dose (15 mg/kg,shown in the left side bar), Isis 405879 significantly (p<0.023) reducedPCSK9 mRNA levels. When administered at a high dose (30 mg/kg, shown inthe right side bar), Isis 405879 also significantly (p<0.011) inhibitedPCSK9 mRNA expression. The results obtained with Isis 405879 weresuperior to those obtained with Isis 395186 (labeled “844419”) and Isis405891 (labeled “844423”). As shown in FIG. 11, Northern analysis ofliver PCSK9 mRNA confirms that 15 mg/kg and 30 mg/kg Isis 405879substantially reduces PCSK9 mRNA levels.

As described in Example 17.1, the reduction of PCSK9 mRNA is expected tocorrelate with a reduction in plasma LDL-C and liver TG. Consistent withthis expectation, Isis 405879 produced a significant (p<0.06) decreasein cyno serum LDL-C after five weeks of administration at 30 mg/kg, asshown in FIG. 12. Isis 405879 additionally produced a significant(p<0.05) increase in the ratio of HDL to LDL following five weeks ofadministration at 30 mg/kg, as shown in FIG. 13.

Generally confirming earlier data obtained in rodents, Isis 405879elicited no differences in serum chemistry, hematology, or organweights, except for a slight increase in spleen weight. Histologyrevealed indicia of only mild inflammation. The advantageousdistribution of Isis 405879 in liver and kidney in the cyno model isdescribed above and shown in FIG. 9.

Example 18: Dose Response Study of Isis 405879 (“844421”) in a HumanPCSK9 Transgenic Mouse Model

BMS-844421 is a human specific PCSK9 antisense oligonucleotide (ASO)designed to downregulate hepatic PCSK9 expression, increase LDL receptoractivity, and promote clearance of LDL and reduction of plasma LDL-Clevels in humans. A human genomic PCSK9 transgenic mouse model wasrecently established. This mouse line expresses hemizygous transgene inthe presence of endogenous mouse PCSK9 (i.e., both genes aresimultaneously expressed). It was found that the transgenic PCSK9 miceresponded to chronic dosing with BMS-844421 and related human PCSK9 ASOswith decreased liver mRNA expression and decreased circulating humanPCSK9 protein levels.

The present study examined the dose response to BMS-844421 in thehemizygous genomic transgenic mouse model on a normal chow diet.Experiments were conducted comparing the effects of BMS-844421 (humanspecific PCSK9 ASO) to those of ISIS 394816 (murine specific PCSK9 ASO)in this model alone and in combination. BMS-844421 exhibiteddose-dependent suppression of liver target mRNA, reflected indose-dependent decreases in plasma human PCSK9 levels. Both liver mRNAand plasma protein gave ED50 values of approximately 15 mg/kg (giventwice weekly) for BMS-844421, while the suppression of plasma PCSK9protein was greater than liver PCSK9 mRNA at each dose in this model.Trends for increased LDLR protein in liver total membrane fraction, anda feedback effect on liver HMGCoA reductase mRNA expression wereobserved, but no changes in plasma lipids were seen for BMS-844421treated hemizygous mice. The combination of ISIS 394816+BMS-844421 (eachat 15 mg/kg) elicted a greater increase in liver LDLR protein thaneither ASO alone, consistent with the proposed mechanism of action andthe dual expression of both human and murine PCSK9 in this model.

18.1. Materials and Methods

The sequences of BMS-844421 and other ASOs tested in this report aregiven in Table 35. The negative control ISIS 141923 is not complementaryto any known gene. The design of the dose response study is described inthe beginning of the Results section (section 18.2. below).

TABLE 35 Nucleotide sequences of ASOs SEQ ID ASO Sequence NO BMS 844421,human 5′-CCTTGGCCACGCCGGCATCC-3′ 248 PCSK9 specific* ISIS 141923,5′-CCTTCCCTGAAGGTTCCTCC-3′ 465 non-targeting ISIS 394816, murine5′-GGGCTCATAGCACATTATCC-3′ 466 PCSK9 specific *BMS-844421 is identicalto ISIS 405879

Parameters evaluated at study termination included hepatic mRNA, hepaticLDLR protein levels, plasma concentrations of total cholesterol, LDL,HDL, triglycerides, transaminases, and organ and body weights. Wholeliver was processed for RNA, and protein, and held for possiblehistological examination and hepatic triglyceride levels. Blood andliver samples were obtained 24 hr after the prior dose. Blood sampleswere obtained from tail vein or retro orbital plexus and EDTA, chilledon ice, and plasma samples were obtained by standard procedures. FormRNA assays, liver samples (30-50 mg each) were placed in eppendorftubes containing 1000 uL of TRIzol (Invitrogen #15596-018) andhomogenized using stainless steel beads on a Tissue Lyser instrument(Qiagen, Valencia, Calif.). Chloroform (200 μL) was added to each tube,the contents were mixed vigorously by hand and tubes were centrifuged at12,000×g for 20 minutes at 4° C. About 300 μL of each supernatant wasremoved and placed in 1.5 mL microcentrifuge tubes. One volume of 70%EtOH was added, the mixtures were vortexed thoroughly and transferred toRNeasy Mini Spin Columns (QIAgen Cat. No. 74106). RNA was purified,treated with DNase and eluted into microcentrifuge tubes followingQIAgen's manufacturer's instructions.

Plasma human PCSK9 levels were assayed using the human specific ELISAwith mAb 4H5 as capture antibody.

18.2. Results

A dose-response study was run in human PCSK9 genomic transgenic mouseline 66 mice on a normal chow diet. Groups of 6 mice (3 males, 3females) received the following treatments for 6 weeks. BMS-844421 wasadministered p.o. at 5, 10, 15, and 30 mg/kg per dose given twice weekly(on Tuesdays and Fridays) for 6 weeks. Controls received saline, and anadditional group received the non-targeting negative control ASO ISIS141923 (30 mg/kg twice weekly). The murine PCSK9 ASO ISIS 394816 wasadministered to another group at 15 mg/kg per dose. The final groupreceived BMS-844421 (15 mg/kg) plus ISIS 394816 (15 mg/kg) combined.

Liver mRNA assayed by qRT-PCR showed that BMS-844421 decreased liverPCSK9 mRNA in a dose dependent manner, with greatest effect at the topdose studied (FIG. 14). The negative control ASO, ISIS 141923, had nosignificant effect on target mRNA, while the murine targeted ASO, ISIS394816, decreased human PCSK9 mRNA by 33%, suggesting partialcross-hybridization to human PCSK9 mRNA.

The ASO effects on plasma human PCSK9 followed a similar pattern asmRNA, while the effects of the higher doses of BMS-844421 resulted ingreater effects, up to 70% suppression at 30 mg/kg/dose (FIG. 15). ISIS394816 suppressed approximately 30% (similar to mRNA effects), whileISIS 141923 had no significant effect.

Plasma human PCSK9 levels exhibited time dependent suppression witheffects at 6 weeks, greater than at 3 wks for all doses of BMS-844421(FIG. 16). At 3 wks dosing, only the highest dose of BMS-844421 showedsignificant suppression. As seen in FIG. 17, data for 6 wks treatmentwith BMS-844421 showed a sigmoidal dose-response curve, with ED50approximately 15 mg/kg/dose (given twice weekly). Plotting the mRNA andprotein data together showed that the two endpoints generally, thoughimperfectly, reflect each other, with protein showing greatersensitivity to suppression (FIG. 18).

Male transgenic mice responded more sensitively to BMS-844421 than thefemale transgenic mice in this study (FIG. 19). The data was not theresult of different plasma baseline levels of human PCSK9 between malesand females, which is human PCSK9=67+/−31 ng/mL in males and 60+/−27ng/mL in females in the predosing samples. The difference in PCSK9protein response for males vs. females was less apparent for livertarget mRNA (FIG. 20).

Levels of endogenous mouse PCSK9 mRNA in liver were decreased by ISIS394816 (and the combination of ISIS 394816 with BMS-844421), but werenot decreased by BMS-844421 only (FIG. 21).

A key cholesterol regulatory gene, HMGCoA reductase (HMGR), is known torespond to sterol feedback in the liver. An assay of endogenous mouseliver HMGR mRNA revealed 45% suppression of HMGR with BMS-844421 as wellas ISIS 394816 (FIG. 22). These findings are consistent withdownregulation of PCSK9 promoting higher levels of LDLR, in turn leadingto increased cholesterol in the regulatory pool in liver cells,resulting in downregulation of HMGR. Similar, though quantitativelylesser effects, were also seen for the sterol pathway genes LDLR mRNAand for SREBP2 mRNA.

LDLR protein levels were assayed in liver samples from the same study.Total hepatic membranes were prepared, and western blots of SDS-PAGEgels were assayed with anti-LDLR antibodies. As a control, transferrinreceptor (TR) protein levels were measured with anti-TR antibodies.Liver samples from the ASO study were pooled from n=3 for thepreparation of the membrane fraction. Pairs of pools for each group wereassayed. Treatment with BMS-844421, and especially the combination ofBMS-844421 with the murine ASO ISIS 394816, led to increases upto >2-fold in levels of LDLR protein, compared to the level of TRprotein (FIG. 23 and FIG. 24).

The ELISA assay uses a monoclonal capture antibody and a polyclonaldetector antibody developed against human recombinant PCSK9 produced inbaculovirus. The recombinant human PCSK9 protein also serves as standardantigen for the ELISA assay. Cynomolgus monkey plasma PCSK9 was readilydetectable using this assay. Human PCSK9 protein in plasma of humanPCSK9 transgenic mouse lines was specifically detected with nocross-reactivity to murine PCSK9.

Despite these pharmacodynamic responses to BMS-844421, hemizygoustransgenic mice did not exhibit changes in plasma LDL-C or totalcholesterol following ASO treatment, including BMS-844421 (Table 36).However, normal chow fed mice have low levels of LDL-C (Table 36). It isplausible that these low LDL-C levels are not readily decreased byincreases in LDLR protein. Another possible explanation is that thehemizygous expression of human PCSK9, i.e., the gene specificallytargeted by BMS-844421, contributes only about ⅓ to ½ of the totalmurine+human PCSK9 in this hemizygous transgenic mouse model.

TABLE 36 Plasma lipids at 6 wk treatment with ASOs in transgenic miceTotal Both Sexes Triglycerides Cholesterol HDL LDL Saline 72 ± 14  108 ±8.2 57 ± 6.4 14 ± 1 Missense 64 ± 9.2 110 ± 8.6 57 ± 7.1 15 ± 1BMS-844421 54 ± 8.6 129 ± 7.3 67 ± 4.6 18 ± 2 (30 mg/kg) BMS-844421 54 ±9.0 126 ± 9.2 65 ± 5.5 18 ± 1 (15 mg/kg) BMS-844421 53 ± 9.4 118 ± 9.361 ± 6.9 17 ± 2 (10 mg/kg) BMS-844421 56 ± 9.7 117 ± 8.5 61 ± 6.1 17 ± 1(5 mg/kg) Mouse ASO 50 ± 7.0 84.2 ± 2.9  45 ± 3.0 11 ± 2 I-394816 (15mg/kg) BMS-844421 + 55 ± 9.3  92 ± 5.3 48 ± 4.6 12 ± 2 I-394816 (15mg/kg each) WT Littermates 60 ± 8.1 111.7 ± 7.2   57 ± 5.3 15 ± 1There were no changes in ALT or AST (not shown), suggesting that noliver abnormalities were present after 6 wks treatment.

18.3. Conclusions

The recently established human genomic PCSK9 transgenic mouse modelresponded to human PCSK9 ASO treatment with robust downregulation ofhuman PCSK9 at both the mRNA and protein levels in liver. Plasma humanPCSK9 was also decreased with the magnitude of decrease somewhat greaterthan liver mRNA suppression, suggesting that plasma PCSK9 levels are agood readout for suppression of the target mRNA. The ED50 for bothendpoints was approximately 15 mg/kg (per dose given twice weekly). Thetransgenic mice did not exhibit significant changes in plasma LDL-C ortotal cholesterol following BMS-844421 or ISIS 394816 treatment, nor forthe combination of the two ASOs. A trend for increasing liver LDLRprotein was observed following treatment with BMS-844421, suggesting afunctional effect consistent with the downregulation of PCSK9 mRNA andprotein. Consistent with this was the downregulation of the sterolresponse gene HMGR in liver.

The limited pharmacodynamic responses in this model appear to be relatedboth to the small LDL window in normal chow diet fed mice, and to theco-expression of the human PCSK9 gene with the endogenous murine PCSK9gene. In the hemizygous mice, human PCSK9 appears to represent onlyabout ⅓ of the total human+murine PCSK9 expressed. Consistent with this,in a previous study the murine specific PCSK9 ASO ISIS 394816administered to human apoB-CETP double transgenic mice produced up to90% suppression of liver PCSK9 mRNA and a corresponding decrease of upto 80% in plasma LDL-C levels.

1. A compound comprising a modified oligonucleotide 12 to 30 linkednucleosides in length and having a nucleobase sequence comprising aportion of at least 8 contiguous nucleobases complementary to anequal-length portion of nucleobases 3543-3569 of SEQ ID NO: 1, andwherein the nucleobase sequence of the modified oligonucleotide is atleast 90% complementary to SEQ ID NO:
 1. 2. The compound of claim 1,wherein the modified oligonucleotide is at least 95% complementary toSEQ ID NO:
 1. 3. The compound of claim 1, wherein the modifiedoligonucleotide is 100% complementary to SEQ ID NO:
 1. 4.-12. (canceled)13. The compound of claim 1, wherein the compound is single-stranded.14. The compound of claim 1, wherein the modified oligonucleotidecomprises at least one internucleoside linkage which is aphosphorothioate internucleoside linkage.
 15. The compound of claim 14,wherein each internucleoside linkage is a phosphorothioateinternucleoside linkage.
 16. The compound of claim 1, wherein themodified oligonucleotide comprises at least one modified sugar.
 17. Thecompound of claim 16, wherein at least one modified sugar is a bicyclicsugar.
 18. The compound of claim 16, wherein at least one modified sugarcomprises a 2′-O-methoxyethyl.
 19. The compound of claim 1, wherein themodified oligonucleotide comprises at least one modified nucleobase. 20.The compound of claim 19, wherein the modified nucleobase is a5-methylcytosine.
 21. The compound of claim 1, wherein the modifiedoligonucleotide comprises: a gap segment consisting of linkeddeoxynucleosides; a 5′ wing segment consisting of linked nucleosides;and a 3′ wing segment consisting of linked nucleosides; wherein the gapsegment is positioned between the 5′ wing segment and the 3′ wingsegment; and wherein each nucleoside of each wing segment comprises amodified sugar.
 22. The compound of claim 21, wherein the modifiedoligonucleotide comprises: a gap segment consisting of ten linkeddeoxynucleosides; a 5′ wing segment consisting of five linkednucleosides; and a 3′ wing segment consisting of five linkednucleosides; wherein the gap segment is positioned between the 5′ wingsegment and the 3′ wing segment, wherein each nucleoside of each wingsegment comprises a 2′-O-methoxyethyl sugar; and wherein eachinternucleoside linkage is a phosphorothioate linkage.
 23. (canceled)24. The compound of claim 2, wherein the modified oligonucleotideconsists of 20 linked nucleosides.
 25. A composition comprising thecompound of claim 1, or a salt thereof and a pharmaceutically acceptablecarrier or diluent.
 26. (canceled)
 27. (canceled)
 28. A methodcomprising administering the compound of claim 1 to an animal having, orat risk of having, a disease associated with PCSK9.
 29. The method ofclaim 28, wherein the animal is a human.
 30. The method of claim 28,wherein administering the compound to the animal slows progressionand/or ameliorates hypercholesterolemia, acute coronary syndrome,polygenic hypercholesterolemia, mixed dyslipidemia, coronary heartdisease, early onset coronary heart disease, type II diabetes, type IIdiabetes with dyslipidemia, hepatic steatosis, non-alcoholicsteatohepatitis, non-alcoholic fatty liver disease,hypertriglyceridemia, hyperfattyacidemia, hyperlipidemia, metabolicsyndrome, atherosclerosis, or improves cardiovascular outcome, or anycombination thereof in the animal.
 31. The method of claim 28 comprisingco-administering the compound and at least one additional therapy.32.-39. (canceled)
 40. The method of claim 28, wherein the administeringis parenteral administration. 41.-281. (canceled)